细胞培养技术五十年演变：从静态二维到智能旋转三维动态培养的综述

1 引言：细胞培养技术的历史跨越

       细胞培养技术作为现代生命科学研究的基石，自20世纪初诞生以来，经历了从简单到复杂、从静态到动态、从二维到三维的波澜壮阔的演进历程。这一演进不仅推动了基础生物学研究的飞速发展，也为药物研发、组织工程和再生医学提供了强大工具。过去五十年间，细胞培养技术经历了四个明显发展阶段，每一个阶段的跃迁都体现了科学家对细胞微环境理解不断深化，也反映了工程技术与生物学的深度融合。

       细胞培养技术的根本挑战在于如何在体外尽可能精确地模拟体内环境。生物体内的细胞生活在复杂的三维结构中，周围是由细胞外基质、邻近细胞和信号分子构成的动态微环境，同时不断接受流体剪切力、机械张力和化学梯度等多种物理化学信号的刺激。传统的二维培养系统虽然操作简便，但与体内环境的差异极大限制了其在预测体内生理响应方面的价值。

       在细胞培养技术的发展历程中，技术驱动和需求牵引两者共同构成了进步的合力。一方面，材料科学、微加工技术和计算机控制的进步为培养系统的创新提供了可能；另一方面，药物筛选对高内涵模型的需求、再生医学对功能性组织构建的追求，都推动了培养技术的革新。特别是在过去二十年间，三维动态培养技术的成熟和普及，极大地改变了体外细胞实验的面貌，使得类器官培养、肿瘤模型构建和个性化医疗等前沿应用成为可能。

        本文将系统回顾半个世纪以来细胞培养技术的演变历程，着重分析从静态二维培养到旋转三维动态培养的技术演进路径，并深入探讨中国科研人员和企业在这一过程中从技术追随者到创新者的角色转变，特别是赛吉生物SARC系列的技术突破与创新价值。

2 早期静态2D培养：技术奠基与局限

       静态二维细胞培养技术的起源可以追溯到20世纪初，1907年Harrison进行的蛙神经组织培养实验普遍被认为是该领域的起点。然而，现代细胞培养技术的真正奠基则归功于1950年代一系列突破性进展，特别是细胞系建立、培养基优化和无菌技术标准化这三个关键领域的发展。1951年，Gey成功分离并培养了著名的人宫颈癌细胞系HeLa，这是第一个可以在体外无限增殖的人类细胞系，为生物医学研究提供了极为宝贵的工具。

       静态2D培养的核心特征在于细胞在平面基质上生长，并暴露于静态培养基中。这种培养模式依赖于几个关键组成部分：培养容器（如培养瓶、培养皿）、合成培养基（包含营养物质、生长因子和激素）、血清（通常为胎牛血清，提供未知生长因子）以及维持恒定理化条件的培养箱。这种技术组合的成功标准化，使得细胞培养从一门艺术转变为一门可重复的科学，极大地推动了细胞生物学、病毒学和癌症研究的发展。在静态2D培养的技术体系中，几种经典方法至今仍被广泛使用。贴壁培养适用于能够在固体基质上粘附并生长的细胞类型，如成纤维细胞、上皮细胞和内皮细胞；悬浮培养则用于非粘附性细胞，如血液细胞和某些癌细胞；原代培养直接从组织分离细胞，保持较高的体内相似性；而传代培养则使细胞系能够长期维持并在实验室间共享。

       静态2D培养技术因其操作简便、成本低廉和可重复性强等优势，迅速成为实验室的标配方法，应用范围极其广泛。在病毒生产领域，2D培养是疫苗生产的关键技术；在癌症研究中，它使科学家能够直接观察癌细胞的分裂和迁移行为；在药物筛选中，它提供了高通量筛选的可行性；在毒性测试中，它是评估化合物细胞毒性的标准模型；而在基础细胞生物学研究中，它为了解细胞基本生命过程提供了窗口。然而，随着研究的深入，静态2D培养的严重局限性也逐渐显现。首先，细胞在二维平面上的生长方式与其在三维组织中的天然状态存在根本差异，这导致细胞极性改变、细胞-细胞连接减少以及细胞-基质相互作用简化。例如，肝细胞在体外迅速失去药物代谢能力，神经元难以建立复杂的神经网络，而肿瘤细胞则无法形成体内的异质性结构。这些现象均表明，二维环境无法提供维持细胞完全分化状态所需的全部信号。

       其次，静态培养系统中的质量传输效率极低，仅依靠分子扩散来提供营养和清除废物。这导致在单层细胞中形成营养梯度和代谢废物积累，特别是在高密度培养时，细胞微环境高度异质，影响实验结果的可靠性和可重复性。此外，静态培养缺乏机械刺激，而生物体内的许多组织（如骨骼、软骨和血管）在正常生理过程中都会经历各种力学信号的影响，这些信号的缺失进一步降低了2D培养细胞的体内相关性。尽管存在这些局限性，静态2D培养作为细胞培养技术发展的基石地位不容置疑。它建立了细胞培养的基本技术框架和标准操作程序，为后续技术演进提供了起点和参照。即使在今天，由于其简单性和低成本，静态2D培养仍然是许多研究实验室的入门技术和常规实验的首选，特别是在需要快速初步筛选的场景中。

3 动态2D培养技术的革新

       面对静态2D培养的固有局限性，科学家们开始探索引入动态培养策略，即在培养过程中通过外力介导培养基流动，以改善细胞微环境。动态2D培养的早期尝试可以追溯到1970年代，当时研究人员开始在搅拌瓶和转瓶中进行大规模细胞培养，主要用于疫苗生产。这些简单的动态系统通过持续混合培养基，减少了营养梯度，提高了细胞产量，标志着细胞培养从静态向动态过渡的重要转折点。动态2D培养的核心优势在于其通过流体运动增强了质量传输效率。在静态培养中，营养物质的供应和代谢废物的移除完全依赖分子扩散，这限制了细胞生长的密度和寿命。而在动态培养中，对流传输成为主导，能够持续为细胞提供新鲜培养基并稀释有害代谢物，从而支持更高密度的细胞生长。此外，流体流动还能在细胞表面产生剪切应力，对于某些细胞类型（如内皮细胞）而言，这种力学刺激是维持其生理功能的重要因素。

       1980年代至1990年代，随着生物反应器技术的进步，动态2D培养进入了快速发展阶段。各种类型的生物反应器被开发出来，包括搅拌罐生物反应器、气升式生物反应器和中空纤维生物反应器。这些系统能够精确控制温度、pH、溶氧和营养浓度等多个环境参数，为细胞提供更稳定、更可控的培养环境。其中，搅拌罐生物反应器通过叶轮搅拌实现培养基均匀混合，特别适用于悬浮细胞的大规模培养；而中空纤维生物反应器则通过半透膜结构模拟毛细血管床，允许小分子物质自由扩散，从而支持高密度细胞培养。

       动态2D培养技术的进步也推动了工业化细胞培养的发展。在单克隆抗体、重组蛋白和疫苗等生物制品的生产中，动态培养系统能够实现从实验室规模（几升）到工业生产规模（几千升）的放大，满足了生物技术产业对一致性和规模的双重要求。例如，用于生产促红细胞生成素(EPO)的细胞培养系统就是动态2D技术成功工业化的典型代表。在科学研究层面，动态2D培养为解决特定生物学问题提供了新的视角。例如，在血管生物学研究中，流动腔系统被广泛应用于研究内皮细胞对流体剪切力的响应，这为了解动脉粥样硬化的发病机制提供了重要线索。在骨骼生物学中，研究人员通过施加机械张力模拟负重状态，研究成骨细胞的分化和骨形成机制。在肿瘤生物学中，动态培养被用于研究循环肿瘤细胞的存活和转移机制。这些应用都表明，力学刺激作为微环境的重要组成部分，在调节细胞行为中发挥着不可替代的作用。

       然而，动态2D培养仍然面临着根本性挑战——细胞还是在二维平面上生长，无法再现组织的三维结构。这意味着，尽管动态2D培养在质量传输和力学刺激方面取得了进步，但它仍然无法完全模拟细胞在体内的真实微环境。细胞在三维组织中的空间排列、梯度分布和细胞-基质相互作用等关键特征在二维系统中仍然缺失。这一认识促使研究人员进一步探索三维培养系统，从而开启了细胞培养技术的新篇章。值得注意的是，在动态2D培养技术的发展过程中，欧美企业凭借其先发优势和技术积累，长期主导着相关设备和耗材的市场。诸如赛默飞世尔、赛多利斯和Eppendorf等跨国公司提供了从实验室到生产级别的全方位解决方案，而中国的生物技术企业在这一领域大多处于追随者角色。这种市场格局在后续的三维动态培养阶段开始发生改变。

4 静态3D培养：维度突破与新挑战

       静态三维培养概念的兴起标志着细胞培养技术演进中的一个范式转变。与旨在改善细胞微环境物理侧面的动态2D培养不同，静态3D培养专注于重建细胞的空间维度，这一转变基于一个日益明确的认知：细胞在三维环境中的行为与其在二维表面上的行为存在根本差异。早在1980年代，一系列经典实验就表明，当乳腺上皮细胞在三维基质中培养时，它们会形成具有极性和分泌功能的腺泡样结构，而在二维培养中则仅呈现单层生长——这一现象凸显了三维环境对细胞功能的重要影响。

       静态3D培养的核心原理是为细胞提供三维支架，模拟体内的细胞外基质(ECM)。这种支架不仅作为物理支持结构，更通过其生化成分、机械性能和拓扑结构主动调节细胞行为，影响细胞的存活、增殖、分化和迁移。与2D培养相比，3D环境更好地保存了细胞的关键特性，如细胞极性、细胞-细胞通讯和细胞-ECM相互作用，这些对维持组织特异性功能至关重要。静态3D培养的实现依赖于多种技术方法，每种方法各有其优势和适用范围。支架基础培养使用天然或合成材料作为支撑结构，是最早发展的3D培养策略之一。天然聚合物支架（如胶原蛋白、Matrigel）生物相容性好但批次间差异大；合成聚合物支架（如聚乳酸-羟基乙酸共聚物PLGA）则具有更好的可控性和可重复性。无支架培养则通过悬滴或强制聚集等方法使细胞自组装成球体，操作简单但不能控制球体的大小和组成。水凝胶培养结合了前两者的优点，通过交联聚合物网络形成高度含水的微环境，既能提供力学支持又允许营养扩散，是目前最有前景的3D培养策略之一。

       静态3D培养在多个研究领域展现出独特价值。在癌症研究中，肿瘤球体模型能够更好地模拟体内的肿瘤结构和药物抵抗性；在干细胞研究中，3D环境有助于干细胞的自我更新和分化，促进类器官的形成；在药物开发中，3D细胞模型显示出比2D模型更好的临床预测性，有望减少药物研发过程中的损耗率；在毒性测试中，3D肝球体和心肌球体分别提供了更可靠的肝毒性和心脏毒性评估模型。尽管静态3D培养在空间结构上更接近体内环境，但它仍然面临物质传输的根本挑战。在三维结构中，营养和氧气的扩散距离增加，中心区域的细胞容易因营养不足和代谢废物积累而死亡，形成坏死核心。这种现象在直径超过200-500微米的球体中尤为明显，限制了三维组织的厚度和复杂性。此外，静态3D培养同样缺乏生理相关的力学刺激，如流体剪切力，而这种力学刺激对于许多组织（尤其是血管和骨骼）的发育和功能至关重要。

       静态3D培养的这些局限性促使研究人员进一步探索将三维结构与动态培养相结合的可能性。同时，在静态3D培养技术的商业化过程中，欧美企业继续保持着领先地位。美国公司Corning的Matrigel和Transwell系统，Lonza的3D细胞培养产品线，以及德国Greiner Bio-One的3D培养板等产品，几乎垄断了全球市场。这种局面直到中国政府对生物技术领域加大投入，以及国内企业如赛吉生物等开始注重自主创新才逐渐改变。静态3D培养代表了细胞培养技术从简单到复杂、从体外到体内模拟的重要过渡阶段。它确立了三维微环境对细胞功能的重要性，为后续动态3D培养技术的发展奠定了概念基础和技术储备。尽管存在局限性，静态3D培养至今仍在许多研究场景中广泛应用，特别是在需要高通量筛选和成本控制的研究中，它提供了在二维和更复杂三维动态模型之间的折衷方案。

5 动态3D培养：流体力学与生物学的结合

       动态3D培养技术的兴起标志着细胞培养领域向着更高生理相关性和复杂性迈出了关键一步。这一技术范式将三维结构的优势与动态培养的质量传输和力学刺激优势相结合，创造出了前所未有的体外模型系统。动态3D培养的理论基础建立在组织工程学、流体力学和细胞生物学的交叉点上，其核心认识是：生物体的细胞不仅生活在三维空间中，还持续暴露于由血液流动、组织变形和间质液流动产生的复杂力学环境中。在动态3D培养系统中，流体运动通过多种机制影响细胞行为。首先，对流传输显著改善了营养物质和氧气的供应，同时有效移除代谢废物，支持更大、更致密的组织构建体存活。其次，流体在细胞表面产生的剪切应力直接调节细胞形态、功能和基因表达。此外，在生物反应器中，悬浮培养物经历的随机运动或定向旋转还能防止细胞沉降和聚集，促进均匀生长。这些优势使得动态3D培养特别适合构建工程化组织、疾病模型和药物测试平台。

       动态3D培养技术的进展与生物反应器系统的创新密不可分。不同类型的生物反应器采用不同策略来创造动态条件。搅拌罐生物反应器通过叶轮搅拌维持细胞悬浮和混合，适合大规模3D培养但可能产生损害性的湍流；灌流生物反应器使培养基连续流过固定的3D支架，提供可控的流体剪切力但可能产生不均匀流动；旋转瓶生物反应器通过容器旋转防止细胞沉降，产生温和混合但规模有限；而回转生物反应器则通过水平轴旋转创造持续自由落体条件，产生低剪切力环境。在这些生物反应器技术中，美国国家航空与宇宙航行局(NASA)开发的旋转壁式生物反应器(Rotating Wall Vessel Bioreactor， RWVB)具有特殊地位-4。1990年，Kleis等人率先研制了一种旋转生物反应器，随后NASA在此基础上改进并开发了RWVB，将其成功应用于组织培养领域-7。RWVB由两个同心圆柱体构成，将细胞与培养液置于内外圆柱体之间，使装置绕纵轴旋转，并通过调节转速使培养物维持悬浮状态-4。该设计形成了一种低剪切力、高物质传递效率并富含溶解氧的液体充盈培养环境。大量研究表明，RWVB能够有效模拟微重力条件下的生物效应，其理论基础在于通过三维空间内持续改变重力矢量，使细胞无法对重力方向产生定向响应——这一机制被称为重力矢量叠加技术。

       NASA最初开发RWVB的目的是在太空飞行中研究微重力对细胞的影响，并保护培养物在发射和着陆期间免受高剪切力伤害-8。但科学家很快意识到，即使在地面实验室，这种系统也能为细胞提供独特的培养环境。RWVB培养物经历的持续自由落体状态使细胞悬浮在培养基中，相互接触但仅暴露于极低的流体剪切力，这惊人地模拟了微重力效应，因此被称为"模拟微重力"培养系统。动态3D培养在组织工程中的应用展示了其强大潜力。在软骨组织工程中，灌流生物反应器被用于培养软骨细胞-支架构建体，流体剪切力促进细胞外基质合成，形成具有机械完整性的组织；在血管工程中，脉动流系统模拟血压的周期性变化，促进内皮细胞和平滑肌细胞形成功能性血管结构；在肝脏组织工程中，3D流动系统支持肝细胞和非实质细胞共培养，维持肝功能标志物的表达和代谢活性；在骨组织工程中，机械刺激（如流体剪切力和循环应变）被证明是诱导成骨细胞分化和矿化的关键因素。

       在动态3D培养技术的发展过程中，欧美科研机构和企业继续保持着技术主导地位。美国公司Synthecon延续NASA的技术路线，商业化生产旋转细胞培养系统(RCCS)，包括STLV（慢转向 lateral vessel）和HARV（高纵横比 vessel）等类型的培养容器。德国Eppendorf和瑞士Infors等生物反应器制造商则提供适用于不同规模和应用的专业发酵罐和生物反应器系统。这些欧美企业的共同特点是注重核心技术研发、系统集成和全球化服务网络，使其产品能够满足从基础研究到工业生产的广泛需求。然而，欧美技术的垄断地位也带来了问题：设备价格昂贵，培养耗材成本高，技术支持在部分地区不足，以及技术方案不一定适合所有地区的实际需求。这些因素为后来中国企业在动态3D培养领域的崛起提供了机会和市场空间。

       动态3D培养技术，特别是旋转壁式生物反应器，代表了细胞培养技术发展中的一个重要里程碑。它不仅在概念上突破了传统培养模式的局限，也为组织工程、疾病建模和药物开发提供了强有力的工具。更重要的是，它建立了一个技术框架，为后续更专业化、更智能化的培养系统——如赛吉生物的SARC系列——奠定了基础。随着对这一技术理念理解的深入和应用经验的积累，细胞培养技术即将迎来新一轮的创新浪潮。

6 旋转3D动态培养的突破性进展

       旋转3D动态培养技术的演进代表了细胞培养领域从简单模拟体内环境到精确重建细胞微环境的重要转变。在这一技术路径中，NASA开发的旋转壁式生物反应器(RWVB)奠定了科学基础，而后续的商业化产品则进一步优化了其设计和功能。旋转细胞培养系统(RCCS)作为RWVB的直接衍生品，由NASA约翰逊航天中心开发，最初目的是在航天飞行期间保护脆弱的组织培养物-8。然而，这一系统提供的低剪切力、高质量传递和模拟微重力的独特环境，很快显示出在地面实验室进行3D细胞生长的明显优势。RCCS的工作原理基于一个简单而精巧的设计：培养容器绕水平轴持续旋转，使培养物保持悬浮状态。根据细胞的种类、性质、数量和培养物的大小调节容器的旋转速度，可使培养物长时间保持悬浮状态-7。这种培养环境形成湍流较少、剪切力低，物质传递效率高，并有丰富的供氧-4。在这种系统中，重力矢量被持续重新定向，使细胞没有足够时间对这种变化作出反应，这被称为重力矢量叠加技术，是模拟微重力效应的理论基础。

       RCCS系统的两种最原始类型的培养容器由Synthecon制造——STLV（慢转向 lateral vessel）和HARV（高纵横比 vessel）。STLV是管状的并且具有中轴气体传输核心，被设计主要用于贴壁细胞培养；HARV则具有盘形培养室，其中氧合膜位于培养容器内壁，被设计主要用于悬浮细胞培养。然而，许多实验表明，它也是贴壁细胞和组织外植体的优良培养容器。随着技术的推广和应用经验的积累，旋转3D动态培养系统显示出在多个领域的独特价值。在组织工程中，它支持复杂3D组织结构形成；在癌症研究中，它能更好地模拟肿瘤微环境；在干细胞生物学中，它促进干细胞分化和类器官形成；在感染性疾病研究中，它提供宿主-病原体相互作用的更真实模型；在药物开发中，它提高了临床前测试的预测价值。这些应用共同证明了旋转3D动态培养在提升体外模型生理相关性方面的巨大潜力。然而，传统的RCCS系统也存在一定局限性：自动化程度低，实时监测能力有限，操作复杂度高，且价格昂贵。这些因素限制了其在资源有限环境中的广泛应用。正是这些挑战，为中国企业实现技术突破和市场切入提供了契机。

6.1 中国技术的崛起：赛吉生物SARC系列的创新

       在这一共同的技术源流下，欧美科研界主要采用基于此原理的RCCS（旋转细胞培养系统），而国内则以苏州赛吉生物的SARC（单轴旋转培养系统）系列为代表-1。无论是RCCS还是SARC，其核心设计理念均源自NASA的RWVB。SARC系列提供了SARC-G通用性旋转3D培养系统以及SARC-P系列灌流旋转3D细胞培养系统等两大系列，继承了旋转模拟微重力的物理机制与低剪切力培养环境-5。

       作为后来者的SARC系统，在继承RCCS所有核心优势的基础上，进一步融合了当代自动化与智能算法技术，实现了系统层面的显著提升-9。SARC-G系列支持多通道独立控制与非灌流培养，SARC-P系列则拓展了连续灌流与长期共培养能力-1。具体而言，SARC系统的后发优势体现在三个方面：自动化与识别能力——系统具备培养容器自动识别功能，可对应加载预设培养程序，减少人工干预误差；智能视觉辅助——部分高端型号集成光学监测与图像分析，支持对类器官形成过程的形态学追踪；算法增强控制——内置剪切力自动计算模型，可根据细胞类型与培养阶段动态优化转速策略，在维持微重力模拟效果的同时，进一步保护细胞免受机械损伤-5。

       SARC系统的技术特点充分体现了中国企业在细胞培养设备领域的创新思路。系统创建了一个无气泡、无机械搅拌、无液体湍流的低剪切力和动态的细胞生长环境-1。通过使用反应容器内部完全充满培养液，同时利用反应容器自带的大面积气体交换膜确保反应容器内外的空气充分交换，SARC系统实现了更接近体内条件的培养环境-9。在微重力效应模拟中，该系统允许用户对目标转速或目标微重力水平进行设置，最大模拟能力可达10⁻³g-1。

6.2 SARC系统的技术进步与国产化意义

       SARC系统的另一重要创新在于其模块化设计和灵活性。SARC-G提供了2通道、4通道以及独立4通道等三种标准规格供用户选择，其中独立4通道可分组独立控制-9。而SARC-P则提供了单通道、双通道等两种标准规格-1。这种多规格设计使研究人员能够根据实验需求和预算灵活选择系统配置，提高了技术的可及性。在培养容器方面，赛吉生物也为SARC系统配套开发了专用容器——SG-RWV旋转壁培养容器和SG-PRV灌流反应容器-1-9。这些容器具有等截面气体交换膜（能够提供更好的气体交换能力）、顶部配有无光学畸变的观测窗，以及可重复灭菌使用的设计-9。这些改进既继承了NASA原始设计的精神，又在实用性和经济性方面进行了优化。

       从早期由SYNTHECON制造的STLV与HARV培养容器，到赛吉生物为SARC系统配套开发的SG-RWV容器——其在气体交换膜设计、观测窗光学性能与可重复灭菌方面的改进——反应容器形态虽不断演进，但其根本的气体交换机制与悬浮培养理念，始终延续自NASA最初确立的RWVB技术框架。SARC系统的成功开发和中国本土化生产具有多重意义。从技术角度看，它证明了中国企业在高端生命科学仪器领域实现创新和突破的能力；从市场角度看，它提供了性能相当但成本更低的替代方案，降低了旋转3D动态培养技术的门槛；从科研支持角度看，它为国内研究人员提供了更便捷的技术支持和更快速的售后响应-1。更重要的是，SARC系统的崛起象征着中国在生物技术领域从技术追随者向技术创新者的角色转变。

       旋转3D动态培养技术的演进，从NASA的RWVB到商业化的RCCS，再到智能化的SARC系统，展示了科学研究与技术创新的有机互动。这一历程既体现了基础科学发现对技术发展的指导作用，也反映了市场需求对技术优化的牵引力量。随着旋转3D动态培养技术的不断成熟和普及，它有望在未来生命科学研究和医学进步中发挥更加重要的作用。

7 细胞培养技术的未来展望

       细胞培养技术历经五十年的演变，从简单的静态2D培养发展到今天高度复杂的旋转3D动态系统，这一进程不仅反映了我们对细胞微环境理解不断深化，也体现了多学科交叉融合在技术创新中的关键作用。展望未来，细胞培养技术将继续向更高生理相关性、更高通量和更高智能化方向发展，同时在个性化医疗、组织工程和药物开发等领域发挥更加重要的作用。在未来五到十年内，我们预期细胞培养技术将出现几个明显发展趋势。

       首先，自动化与智能化将成为标准功能。正如赛吉生物SARC系统所展示的，培养容器自动识别、实时剪切力计算和自适应控制等功能将从高端配置逐步普及到基础型号中-1。人工智能算法的引入将使得系统能够根据实时监测的细胞状态自动调整培养参数，实现真正意义上的智能培养。同时，多模态集成传感技术将允许系统同时追踪多种细胞行为指标，为研究人员提供更全面的培养物状态信息。

       其次，微型化与高通量化并行发展。一方面，微流控技术与3D培养的结合将使得在芯片上构建微型器官(organ-on-a-chip)成为可能，这种系统能够精确控制微流体和化学梯度，更好地模拟体内器官的微观结构和发展过程。另一方面，适应药物开发需求的高通量3D培养系统也将不断进步，使研究人员能够在合理时间内筛选大量化合物，加速药物发现进程。这类系统很可能结合液体处理机器人和自动化成像系统，实现从培养到分析的全流程自动化。

       第三，多器官系统的构建将成为现实。目前的研究主要集中在单一器官模型，而未来的技术将致力于连接不同的器官芯片，形成人体-on-a-chip系统，更好地模拟全身性的药物代谢和毒性反应。这种系统将整合肝脏、心脏、大脑和肾脏等多个器官模型，通过微流体网络相互连接，提供比传统动物模型更准确的人体反应预测。这一方向的技术进步将极大改变药物开发流程，可能减少对动物试验的依赖，提高药物研发效率。

在个性化医疗领域，细胞培养技术也将扮演越来越重要的角色。结合患者特异性诱导多能干细胞(iPSCs)和先进的3D动态培养系统，研究人员将能够建立患者特异性疾病模型，用于个性化药物筛选和治疗方案优化。特别是在肿瘤学领域，利用患者肿瘤细胞快速构建的3D肿瘤模型，能够在治疗开始前测试不同药物组合的效果，为精准医疗提供强大工具。这类应用将推动细胞培养技术从基础研究工具向临床决策辅助手段转变。

中国企业在未来细胞培养技术发展中的角色值得特别关注。以赛吉生物为代表的中国生物技术企业，通过SARC系列的成功开发，展示了结合本土需求与全球视野的创新模式-1-5-9。这些企业既理解国内科研人员的具体需求和工作流程，又能够吸收国际先进技术理念，开发出具有竞争力的产品。随着中国对生物技术领域的持续投入和市场需求的不断扩大，我们有理由相信，将会有更多中国企业从技术追随者转变为技术引领者，在全球细胞培养设备市场中占据重要地位。

       中国企业的崛起也将促进全球细胞培养技术的多元化发展和可及性提升。通过提供性能相当但成本更低的替代方案，中国企业使更多资源有限的实验室能够使用先进的3D动态培养技术，从而推动全球科学研究的进步。同时，针对特定区域性疾病（如某些在亚洲高发的疾病）开发的专用培养系统，也将丰富全球研究工具的选择，为解决特定健康问题提供支持。细胞培养技术的未来充满希望，但也面临挑战。如何更好地模拟免疫系统在组织发育和疾病中的作用，如何构建血管化的组织工程构建体，如何平衡系统的复杂性和可用性，这些都是需要继续探索的方向。但无论如何，从静态2D到旋转3D动态培养的技术演进路径已经为我们指明了前进的方向——更加尊重细胞的生物学特性，更加精细地模拟体内微环境，更加智能地控制系统运行。

8 写在最后

       细胞培养技术五十年的演变历程是一段从简单到复杂、从静态到动态、从二维到三维的螺旋式上升过程。这一历程不仅见证了技术本身的进步，也反映了科学研究范式的转变——从试图简化系统以识别基本规律，到努力构建复杂系统以更好地模拟现实世界。在细胞培养技术的发展道路上，几个关键转变尤为明显：从静态单层培养到动态悬浮培养的转变改善了质量传输效率；从二维平台到三维支架的转变重建了细胞空间环境；从均质系统到异质共培养的转变恢复了细胞-细胞相互作用；从固定参数到自适应控制的转变实现了培养过程的优化。这些转变共同指向一个目标：在体外创造尽可能接近体内条件的微环境，使细胞能够在培养皿中展现出其在生物体内的真实行为。

       回顾这一技术演进史，我们也能清晰看到中国在生命科学仪器领域从缺席者到参与者再到创新者的角色转变。从早期完全依赖进口设备，到后来仿制改进，再到今天开发出具有自主知识产权和国际竞争力的产品，如赛吉生物的SARC系列，这一转变不仅体现了中国科技实力的提升，也预示着全球生命科学仪器市场格局的潜在变化。细胞培养技术的进步永远改变了生物学研究的方式。它使科学家能够提出和回答更加复杂的问题，推动了从分子细胞生物学到系统生物学的范式转变。在未来，随着技术的进一步成熟，我们有望看到细胞培养在再生医学、疾病建模和个性化医疗等领域发挥更加重要的作用，最终实现从实验室研究到临床应用的直接转化。

       正如NASA的RWVB意外地为地面细胞培养技术开辟了新路径，细胞培养技术的未来也可能来自于我们今天难以预见的突破和创新。但无论如何，对生命基本规律的尊重和对科学探索的执着，将继续引领这一领域向前发展，为人类健康和科学进步做出新的贡献。

参考文献

SARC-P灌流旋转细胞培养系统. Instrument.com.cn. https://www.instrument.com.cn/show/C625282.html

[未经美国FDA批准或批准的设备试用] (SARCA). ClinicalTrials.gov. https://ichgcp.net/zh/clinical-trials-registry/NCT05339139

Kleis S.J., Schreck S., Nerem R.M. A viscous pump bioreactor. Biotechnology and Bioengineering. 1990. https://www.semanticscholar.org/paper/A-viscous-pump-bioreactor-Kleis-Schreck/8091c2dfac9c9df611e647432a7358690d022f7a

孔德胜, 胡龙虎. 微重力旋转细胞培养的研究及应用进展. 航空航天医学杂志. https://www.sinomed.ac.cn/article.do?ui=2019179358

自动细胞灌流培养系统SARC-P. Instrument.com.cn. https://www.instrument.com.cn/show/C625293.html

萧山经济技术开发区企安心服务枢纽平台. http://xetz.xsnet.cn/index/consult/content?id=35215

微重力旋转细胞培养的研究及应用进展. 万方医学网. https://med.wanfangdata.com.cn/Paper/Detail/PeriodicalPaper_hkhtyy200911001

McCarthy B.E. Past, Present, and Future of NASA Research into Microbes in Space. Arizona State University. https://keep.lib.asu.edu/items/135188?_format=mods

SARC-G通用旋转细胞培养系统. Instrument.com.cn. https://www.instrument.com.cn/show/C625283.html

The Fifty-Year Evolution of Cell Culture Technology: From Static 2D to Intelligent Rotating 3D Dynamic Culture

1 Introduction: The Historical Leap of Cell Culture Technology

As the cornerstone of modern life science research, cell culture technology has undergone a spectacular evolution from simple to complex, from static to dynamic, and from two-dimensional to three-dimensional since its inception in the early 20th century. This evolution has not only driven the rapid development of basic biological research but also provided powerful tools for drug development, tissue engineering, and regenerative medicine. Over the past fifty years, cell culture technology has gone through four distinct developmental stages, with each transitional phase reflecting scientists' deepening understanding of the cellular microenvironment and the deep integration of engineering technology and biology.

The fundamental challenge of cell culture technology lies in simulating the in vivo environment as accurately as possible in vitro. Cells in living organisms reside in complex three-dimensional structures, surrounded by a dynamic microenvironment composed of extracellular matrix, adjacent cells, and signaling molecules, while continuously receiving various physical and chemical signals such as fluid shear stress, mechanical tension, and chemical gradients. Although traditional two-dimensional culture systems are easy to operate, their significant differences from the in vivo environment greatly limit their value in predicting in vivo physiological responses.

In the development of cell culture technology, both technology-driven and demand-pulled factors have jointly contributed to progress. On one hand, advances in materials science, microfabrication technology, and computer control have made innovations in culture systems possible; on the other hand, the demand for high-content models in drug screening, the pursuit of functional tissue construction in regenerative medicine, and research on cell behavior in microgravity environments have all driven technological innovations. Particularly over the past two decades, the maturation and popularization of three-dimensional dynamic culture technologies have dramatically changed the landscape of in vitro cell experiments, enabling cutting-edge applications such as organoid culture, tumor model construction, and personalized medicine.

This article systematically reviews the evolution of cell culture technology over the past half-century, focusing on the technological evolution path from static two-dimensional culture to rotating three-dimensional dynamic culture, and exploring in depth the role transition of Chinese researchers and enterprises from technology followers to innovators, especially the technical breakthroughs and innovative value of Sagebio's SARC series.

2 Early Static 2D Culture: Technological Foundation and Limitations

The origins of static two-dimensional cell culture technology can be traced back to the early 20th century, with Harrison's 1907 experiment on frog nerve tissue culture widely regarded as the starting point of the field. However, the true foundation of modern cell culture technology is credited to a series of breakthrough advances in the 1950s, particularly in three key areas: cell line establishment, culture medium optimization, and standardization of aseptic techniques. In 1951, Gey successfully isolated and cultured the famous human cervical cancer cell line HeLa, the first human cell line capable of unlimited proliferation in vitro, providing an extremely valuable tool for biomedical research.

The core characteristics of static 2D culture lie in cell growth on a planar substrate while exposed to static culture medium. This culture model relies on several key components: culture vessels (such as culture flasks and petri dishes), synthetic media (containing nutrients, growth factors, and hormones), serum (typically fetal bovine serum, providing unknown growth factors), and incubators that maintain constant physicochemical conditions. The successful standardization of this technological combination transformed cell culture from an art to a reproducible science, greatly advancing the development of cell biology, virology, and cancer research.

In the technical system of static 2D culture, several classical methods are still widely used today. Adherent culture is suitable for cell types that can adhere and grow on solid substrates, such as fibroblasts, epithelial cells, and endothelial cells; suspension culture is used for non-adherent cells, such as blood cells and certain cancer cells; primary culture directly isolates cells from tissues, maintaining high in vivo similarity; while subculture enables cell lines to be maintained long-term and shared between laboratories.

Static 2D culture technology quickly became the standard laboratory method due to its advantages of easy operation, low cost, and high reproducibility, with an extremely wide range of applications. In the field of virus production, 2D culture is a key technology for vaccine production; in cancer research, it enables scientists to directly observe cancer cell division and migration behavior; in drug screening, it provides feasibility for high-throughput screening; in toxicity testing, it is the standard model for assessing compound cytotoxicity; and in basic cell biology research, it provides a window into understanding fundamental cellular life processes.

However, as research deepened, the serious limitations of static 2D culture gradually became apparent. First, the growth of cells on two-dimensional surfaces fundamentally differs from their natural state in three-dimensional tissues, leading to altered polarity, reduced cell-cell connections, and simplified cell-matrix interactions. For example, liver cells rapidly lose drug metabolism capacity in vitro, neurons struggle to establish complex neural networks, and tumor cells cannot form heterogeneous structures found in vivo. These phenomena indicate that two-dimensional environments cannot provide all the signals necessary to maintain fully differentiated cell states.

Secondly, mass transfer efficiency in static culture systems is extremely low, relying solely on molecular diffusion to provide nutrients and remove waste products. This leads to the formation of nutrient gradients and metabolic waste accumulation in monolayer cultures, particularly in high-density cultures where the cellular microenvironment becomes highly heterogeneous, affecting the reliability and reproducibility of experimental results. Additionally, static culture lacks mechanical stimulation, while many tissues in living organisms (especially bones, cartilage, and blood vessels) experience various mechanical signals during normal physiological processes. The absence of these signals further reduces the in vivo relevance of 2D cultured cells.

Despite these limitations, the cornerstone status of static 2D culture in the development of cell culture technology is undeniable. It established the basic technical framework and standard operating procedures for cell culture, providing a starting point and reference for subsequent technological evolution. Even today, due to its simplicity and low cost, static 2D culture remains the introductory technique and first choice for routine experiments in many research laboratories, particularly in scenarios requiring rapid preliminary screening.

3 The Innovation of Dynamic 2D Culture Technology

Faced with the inherent limitations of static 2D culture, scientists began exploring dynamic culture strategies, that is, introducing external forces to mediate fluid flow during the culture process to improve the cellular microenvironment. Early attempts at dynamic 2D culture date back to the 1970s, when researchers began large-scale cell culture in spinner flasks and roller bottles, mainly for vaccine production. These simple dynamic systems improved nutrient gradients and increased cell yield through continuous mixing of culture medium, marking an important turning point in the transition from static to dynamic cell culture.

The core advantage of dynamic 2D culture lies in its enhancement of mass transfer efficiency through fluid motion. In static culture, the supply of nutrients and removal of metabolic wastes rely entirely on molecular diffusion, which limits cell growth density and longevity. In dynamic culture, convective transport dominates, enabling continuous provision of fresh medium to cells while diluting harmful metabolites, thereby supporting higher density cell growth. Additionally, fluid flow can generate shear stress on cell surfaces, which for certain cell types (such as endothelial cells) is an important factor in maintaining their physiological functions.

From the 1980s to the 1990s, with advances in bioreactor technology, dynamic 2D culture entered a stage of rapid development. Various types of bioreactors were developed, including stirred-tank bioreactors, airlift bioreactors, and hollow fiber bioreactors. These systems could precisely control multiple environmental parameters such as temperature, pH, dissolved oxygen, and nutrient concentration, providing cells with a more stable and controllable culture environment. Among these, stirred-tank bioreactors achieve uniform mixing of culture medium through impeller agitation, making them particularly suitable for large-scale suspension cell culture; while hollow fiber bioreactors simulate capillary beds through semi-permeable membrane structures, allowing free diffusion of small molecules and thus supporting high-density cell culture.

Advances in dynamic 2D culture technology also promoted the development of industrial cell culture. In the production of biologics such as monoclonal antibodies, recombinant proteins, and vaccines, dynamic culture systems enabled scaling from laboratory scale (a few liters) to industrial production scale (thousands of liters), meeting the dual requirements of consistency and scale in the biotechnology industry. For example, the cell culture system used to produce erythropoietin (EPO) is a typical representative of the successful industrialization of dynamic 2D technology.

At the scientific research level, dynamic 2D culture provided new perspectives for solving specific biological problems. For instance, in vascular biology research, flow chamber systems are widely used to study endothelial cell responses to fluid shear stress, providing important clues for understanding the pathogenesis of atherosclerosis. In bone biology, researchers simulate loading conditions by applying mechanical tension to study osteoblast differentiation and bone formation mechanisms. In tumor biology, dynamic culture is used to study the survival and metastasis mechanisms of circulating tumor cells. These applications demonstrate that mechanical stimulation, as an important component of the microenvironment, plays an irreplaceable role in regulating cell behavior.

However, dynamic 2D culture still faces a fundamental challenge—cells still grow on two-dimensional surfaces, unable to reproduce the three-dimensional structure of tissues. This means that although dynamic 2D culture has made progress in mass transfer and mechanical stimulation, it still cannot fully simulate the real microenvironment of cells in vivo. Key characteristics of cells in three-dimensional tissues, such as spatial arrangement, gradient distribution, and cell-matrix interactions, are still missing in two-dimensional systems. This realization prompted researchers to further explore three-dimensional culture systems, thus opening a new chapter in cell culture technology.

It is worth noting that during the development of dynamic 2D culture technology, European and American enterprises, with their first-mover advantages and technological accumulation, long dominated the market for related equipment and consumables. Multinational corporations such as Thermo Fisher, Sartorius, and Eppendorf provided comprehensive solutions from laboratory to production level, while Chinese biotechnology companies mostly played follower roles in this field. This market pattern began to change in the subsequent three-dimensional dynamic culture stage.

4 Static 3D Culture: Dimensional Breakthrough and New Challenges

The rise of static three-dimensional culture concepts marks a paradigm shift in the evolution of cell culture technology. Unlike dynamic 2D culture that focuses on improving the physical aspects of the cellular microenvironment, static 3D culture focuses on reconstructing the spatial dimension of cells. This shift is based on the increasingly clear recognition that cell behavior in three-dimensional environments is fundamentally different from their behavior on two-dimensional surfaces. As early as the 1980s, a series of classic experiments showed that when mammary epithelial cells were cultured in three-dimensional matrices, they formed acinar-like structures with polarity and secretory functions, while in two-dimensional culture they only showed monolayer growth—a phenomenon highlighting the importance of the three-dimensional environment for cell function.

The core principle of static 3D culture is to provide cells with three-dimensional scaffolds that simulate the in vivo extracellular matrix (ECM). These scaffolds not only serve as physical support structures but also actively regulate cell behavior through their biochemical composition, mechanical properties, and topological structure, affecting cell survival, proliferation, differentiation, and migration. Compared with 2D culture, 3D environments better preserve key cell characteristics, such as cell polarity, cell-cell communication, and cell-ECM interactions, which are crucial for maintaining tissue-specific functions.

Static 3D culture is achieved through various technical methods, each with its own advantages and applicable scope. Scaffold-based culture uses natural or synthetic materials as support structures and is one of the earliest developed 3D culture strategies. Natural polymer scaffolds (such as collagen, Matrigel) have good biocompatibility but large batch-to-batch variations; synthetic polymer scaffolds (such as polylactic-co-glycolic acid PLGA) have better controllability and reproducibility. Scaffold-free culture enables cells to self-assemble into spheroids through methods such as hanging drops or forced aggregation, with simple operation but inability to control spheroid size and composition. Hydrogel culture combines the advantages of the previous two, forming a highly hydrated microenvironment through cross-linked polymer networks, which can provide mechanical support while allowing nutrient diffusion, and is currently one of the most promising 3D culture strategies.

Static 3D culture has demonstrated unique value in multiple research fields. In cancer research, tumor spheroid models can better simulate in vivo tumor structure and drug resistance; in stem cell research, 3D environments help with stem cell self-renewal and differentiation, promoting organoid formation; in drug development, 3D cell models show better clinical predictability than 2D models, potentially reducing attrition rates in drug development processes; in toxicity testing, 3D liver spheroids and cardiac spheroids provide more reliable models for hepatotoxicity and cardiotoxicity assessment, respectively.

Although static 3D culture is closer to the in vivo environment in terms of spatial structure, it still faces the fundamental challenge of mass transfer. In three-dimensional structures, the diffusion distance for nutrients and oxygen increases, making cells in central areas prone to death due to nutrient deficiency and metabolic waste accumulation, forming necrotic cores. This phenomenon is particularly evident in spheroids with diameters exceeding 200-500 micrometers, limiting the thickness and complexity of three-dimensional tissues. Additionally, static 3D culture also lacks physiologically relevant mechanical stimulation, such as fluid shear stress, which is crucial for the development and function of many tissues (especially blood vessels and bones).

These limitations of static 3D culture prompted researchers to further explore the possibility of combining three-dimensional structure with dynamic culture. Meanwhile, in the commercialization process of static 3D culture technology, European and American companies continued to maintain their leading position. Products from American company Corning's Matrigel and Transwell systems, Lonza's 3D cell culture product line, and Germany's Greiner Bio-One's 3D culture plates almost monopolized the global market. This situation only began to change gradually after the Chinese government increased investment in the biotechnology field and domestic enterprises such as Sagebio began to focus on independent innovation.

Static 3D culture represents an important transitional stage in cell culture technology from simple to complex, from in vitro to in vivo simulation. It established the importance of the three-dimensional microenvironment for cell function, laying the conceptual foundation and technical reserve for subsequent dynamic 3D culture technology development. Despite its limitations, static 3D culture is still widely used in many research scenarios today, especially in studies requiring high-throughput screening and cost control, where it provides a compromise between two-dimensional and more complex three-dimensional dynamic models.

5 Dynamic 3D Culture: The Combination of Fluid Mechanics and Biology

The rise of dynamic 3D culture technology marks a critical step in the cell culture field toward higher physiological relevance and complexity. This technological paradigm combines the advantages of three-dimensional structure with the mass transfer and mechanical stimulation advantages of dynamic culture, creating unprecedented in vitro model systems. The theoretical foundation of dynamic 3D culture is built on the intersection of tissue engineering, fluid mechanics, and cell biology, with its core recognition that cells in living organisms not only live in three-dimensional space but are also continuously exposed to complex mechanical environments generated by blood flow, tissue deformation, and interstitial fluid flow.

In dynamic 3D culture systems, fluid motion affects cell behavior through multiple mechanisms. First, convective transport significantly improves the supply of nutrients and oxygen while effectively removing metabolic waste, supporting the survival of larger and denser tissue constructs. Second, shear stress generated by fluid on cell surfaces directly regulates cell morphology, function, and gene expression. Additionally, the random motion or directional rotation experienced by suspended cultures in bioreactors can prevent cell sedimentation and aggregation, promoting uniform growth. These advantages make dynamic 3D culture particularly suitable for constructing engineered tissues, disease models, and drug testing platforms.

Advances in dynamic 3D culture technology are inseparable from innovations in bioreactor systems. Different types of bioreactors employ different strategies to create dynamic conditions. Stirred-tank bioreactors maintain cell suspension and mixing through impeller agitation, suitable for large-scale 3D culture but potentially generating damaging turbulence; perfusion bioreactors allow continuous flow of culture medium through fixed 3D scaffolds, providing controllable fluid shear stress but potentially producing uneven flow; rotating bottle bioreactors prevent cell sedimentation through container rotation, producing gentle mixing but with limited scale; while rotating wall bioreactors create continuous free-fall conditions through horizontal axis rotation, producing low shear stress environments.

Among these bioreactor technologies, the Rotating Wall Vessel Bioreactor (RWVB) developed by the National Aeronautics and Space Administration (NASA) holds a special position-4. In 1990, Kleis and others first developed a rotating bioreactor, which NASA subsequently improved upon to develop the RWVB, successfully applying it to the field of tissue culture-7. The RWVB consists of two concentric cylinders, with cells and culture fluid placed between the inner and outer cylinders, and the device rotates around the longitudinal axis, with the rotation speed adjusted according to the type, nature, and quantity of cells and the size of the culture to maintain the culture in suspension-4. This design creates a liquid-filled culture environment with low shear stress, high mass transfer efficiency, and rich dissolved oxygen. Numerous studies have shown that the RWVB can effectively simulate biological effects under microgravity conditions, with its theoretical basis being that by continuously changing the gravity vector in three-dimensional space, cells cannot respond to this change in time—a mechanism known as the gravity vector superposition technique.

NASA initially developed the RWVB to study the effects of microgravity on cells during space flight and to protect cultures from high shear stress damage during launch and landing-8. But scientists quickly realized that even in ground laboratories, this system could provide a unique culture environment for cells. The continuous free-fall state experienced by RWVB cultures keeps cells suspended in the culture medium, contacting each other but exposed to extremely low fluid shear stress, which surprisingly simulates microgravity effects, hence being called a "simulated microgravity" culture system.

The application of dynamic 3D culture in tissue engineering demonstrates its strong potential. In cartilage tissue engineering, perfusion bioreactors are used to culture chondrocyte-scaffold constructs, with fluid shear stress promoting extracellular matrix synthesis and forming tissues with mechanical integrity; in vascular engineering, pulsatile flow systems simulate cyclic blood pressure changes, promoting the formation of functional vascular structures by endothelial cells and smooth muscle cells; in liver tissue engineering, 3D flow systems support the co-culture of hepatocytes and non-parenchymal cells, maintaining the expression of liver function markers and metabolic activity; in bone tissue engineering, mechanical stimulation (such as fluid shear stress and cyclic strain) has been shown to be a key factor inducing osteoblast differentiation and mineralization.

In the development of dynamic 3D culture technology, European and American research institutions and companies continued to maintain their technological dominance. The American company Synthecon continued NASA's technical route, commercially producing the Rotating Cell Culture System (RCCS), including culture vessels such as STLV (slow turning lateral vessel) and HARV (high aspect ratio vessel). Bioreactor manufacturers such as Germany's Eppendorf and Switzerland's Infors provide specialized fermenters and bioreactor systems suitable for different scales and applications. These European and American companies share common characteristics of focusing on core technology research and development, system integration, and global service networks, enabling their products to meet a wide range of needs from basic research to industrial production.

However, the monopoly position of European and American technology also brought problems: expensive equipment, high consumable costs, insufficient technical support in some regions, and technical solutions not necessarily suitable for the actual needs of all regions. These factors provided opportunities and market space for the later rise of Chinese enterprises in the field of dynamic 3D culture.

Dynamic 3D culture technology, particularly the rotating wall vessel bioreactor, represents an important milestone in the development of cell culture technology. It not only conceptually broke through the limitations of traditional culture models but also provided powerful tools for tissue engineering, disease modeling, and drug development. More importantly, it established a technical framework for subsequent more specialized and intelligent culture systems—such as Sagebio's SARC series. With the deepening understanding of this technical concept and the accumulation of application experience, cell culture technology is about to usher in a new wave of innovation.

6 Breakthrough Progress in Rotating 3D Dynamic Culture

The evolution of rotating 3D dynamic culture technology represents an important shift in the cell culture field from simply simulating the in vivo environment to accurately reconstructing the cellular microenvironment. In this technological path, the rotating wall vessel bioreactor (RWVB) developed by NASA laid the scientific foundation, while subsequent commercial products further optimized its design and function. The Rotating Cell Culture System (RCCS), as a direct derivative of the RWVB, was developed by NASA's Johnson Space Center, initially aimed at protecting fragile tissue cultures during space flight-8. However, the unique environment of low shear stress, high mass transfer, and simulated microgravity provided by this system quickly demonstrated significant advantages for 3D cell growth in ground laboratories.

The working principle of RCCS is based on a simple yet elegant design: the culture vessel rotates continuously around the horizontal axis, keeping the culture in suspension. Adjusting the rotation speed of the container according to the type, nature, quantity of cells, and the size of the culture allows the culture to remain suspended for extended periods-7. This culture environment produces less turbulence, low shear stress, high mass transfer efficiency, and abundant oxygen supply-4. In such systems, the gravity vector is continuously reoriented, preventing cells from having enough time to respond to this change, which is called the gravity vector superposition technique, the theoretical basis for simulating microgravity effects.

The two most original types of culture vessels for the RCCS system were manufactured by Synthecon—STLV (slow turning lateral vessel) and HARV (high aspect ratio vessel). The STLV is tubular and has a central gas transfer core, designed mainly for adherent cell culture; the HARV has a disk-shaped culture chamber with an oxygenation membrane located on the inner wall of the culture vessel, designed mainly for suspension cell culture. However, many experiments have shown that it is also an excellent culture vessel for adherent cells and tissue explants.

With the promotion of technology and the accumulation of application experience, rotating 3D dynamic culture systems have demonstrated unique value in multiple fields. In tissue engineering, it supports the formation of complex 3D tissue structures; in cancer research, it can better simulate the tumor microenvironment; in stem cell biology, it promotes stem cell differentiation and organoid formation; in infectious disease research, it provides more realistic models of host-pathogen interactions; in drug development, it improves the predictive value of preclinical testing. These applications collectively demonstrate the great potential of rotating 3D dynamic culture in enhancing the physiological relevance of in vitro models.

However, traditional RCCS systems also have certain limitations: low automation, limited real-time monitoring capability, high operational complexity, and high cost. These factors limited their widespread application in resource-limited environments. It was these challenges that provided opportunities for Chinese enterprises to achieve technological breakthroughs and market entry.

6.1 The Rise of Chinese Technology: Innovation of Sagebio's SARC Series

In this common technological origin, the European and American research community mainly adopted the RCCS (Rotating Cell Culture System) based on this principle, while in China, the SARC (Single Axis Rotary Culture) series from Suzhou Sagebio became the representative-1. Whether RCCS or SARC, their core design concepts originate from NASA's RWVB. The SARC series provides two major series: the SARC-G universal rotating 3D culture system and the SARC-P series perfusion rotating 3D cell culture system, inheriting the physical mechanism of rotating simulated microgravity and the low shear stress culture environment-5.

As a latercomer, the SARC system, while inheriting all the core advantages of RCCS, further integrated contemporary automation and intelligent algorithm technology, achieving significant improvements at the system level-9. The SARC-G series supports multi-channel independent control and non-perfusion culture, while the SARC-P series expands continuous perfusion and long-term co-culture capabilities-1. Specifically, the latecomer advantages of the SARC system are reflected in three aspects: automation and identification capability—the system has automatic culture vessel identification function, which can load preset culture programs accordingly, reducing human intervention errors; intelligent visual assistance—some high-end models integrate optical monitoring and image analysis, supporting morphological tracking of organoid formation processes; algorithm-enhanced control—built-in automatic shear force calculation models can dynamically optimize speed strategies according to cell type and culture stage, while maintaining simulated microgravity effects, further protecting cells from mechanical damage-5.

The technical characteristics of the SARC system fully reflect the innovative thinking of Chinese enterprises in the field of cell culture equipment. The system creates a bubble-free, mechanical stirring-free, liquid turbulence-free low shear stress and dynamic cell growth environment-1. By completely filling the reaction vessel with culture fluid and using the large-area gas exchange membrane that comes with the reaction vessel to ensure full air exchange inside and outside the reaction vessel, the SARC system achieves a culture environment closer to in vivo conditions-9. In microgravity effect simulation, this system allows users to set target speed or target microgravity level, with maximum simulation capability up to 10⁻³g-1.

6.2 Technical Progress of SARC System and Significance of Localization

Another important innovation of the SARC system lies in its modular design and flexibility. The SARC-G provides three standard specifications for users to choose from: 2-channel, 4-channel, and independent 4-channel, of which the independent 4-channel can be controlled independently in groups-9. The SARC-P provides two standard specifications: single-channel and dual-channel-1. This multi-specification design enables researchers to flexibly select system configurations according to experimental needs and budget, improving the accessibility of the technology.

In terms of culture vessels, Sagebio has also developed specialized vessels for the SARC system—SG-RWV rotating wall culture vessels and SG-PRV perfusion reaction vessels-1-9. These vessels feature equal-cross-section gas exchange membranes (providing better gas exchange capability), observation windows without optical distortion on the top, and reusable sterilization design-9. These improvements both inherit the spirit of NASA's original design and optimize practicality and economy.

From the early STLV and HARV culture vessels manufactured by SYNTHECON, to the SG-RWV vessels developed by Sagebio for the SARC system—their improvements in gas exchange membrane design, observation window optical performance, and reusable sterilization—although the form of reaction vessels has evolved, their fundamental gas exchange mechanism and suspension culture concept have always continued the technical framework of the RWVB established by NASA.

The successful development and localized production of the SARC system in China have multiple significances. From a technical perspective, it demonstrates the ability of Chinese enterprises to achieve innovation and breakthroughs in the field of high-end life science instruments; from a market perspective, it provides equivalent performance but lower-cost alternatives, lowering the barrier to rotating 3D dynamic culture technology; from a research support perspective, it provides domestic researchers with more convenient technical support and faster after-sales response-1. More importantly, the rise of the SARC system symbolizes the role transition of China in the biotechnology field from technology follower to technology innovator.

The evolution of rotating 3D dynamic culture technology, from NASA's RWVB to commercialized RCCS, and then to intelligent SARC systems, demonstrates the organic interaction between scientific research and technological innovation. This process reflects both the guiding role of basic scientific discoveries on technological development and the pulling force of market demand on technological optimization. With the continuous maturation and popularization of rotating 3D dynamic culture technology, it is expected to play a more important role in future life science research and medical progress.

7 Future Prospects of Cell Culture Technology

After fifty years of evolution, from simple static 2D culture to today's highly complex rotating 3D dynamic systems, the development of cell culture technology not only reflects our deepening understanding of the cellular microenvironment but also embodies the key role of multidisciplinary integration in technological innovation. Looking forward, cell culture technology will continue to develop toward higher physiological relevance, higher throughput, and higher intelligence, while playing an increasingly important role in personalized medicine, tissue engineering, and drug development.

In the next five to ten years, we expect several clear development trends in cell culture technology. First, automation and intelligence will become standard features. As demonstrated by Sagebio's SARC system, functions such as automatic culture vessel identification, real-time shear force calculation, and adaptive control will gradually spread from high-end configurations to basic models-1. The introduction of artificial intelligence algorithms will enable systems to automatically adjust culture parameters based on real-time monitored cell states, achieving true intelligent culture. Meanwhile, multi-modal integrated sensing technology will allow systems to simultaneously track multiple cell behavior indicators, providing researchers with more comprehensive information on culture status.

Second, miniaturization and high-throughput will develop in parallel. On one hand, the combination of microfluidic technology and 3D culture will make it possible to build miniature organs on chips (organ-on-a-chip). Such systems can precisely control microfluids and chemical gradients, better simulating the microstructure and development process of in vivo organs. On the other hand, high-throughput 3D culture systems adapted to drug development needs will also continue to advance, enabling researchers to screen large numbers of compounds within a reasonable time frame, accelerating the drug discovery process. Such systems are likely to combine liquid handling robots and automated imaging systems to achieve full-process automation from culture to analysis.

Third, the construction of multi-organ systems will become a reality. Current research mainly focuses on single organ models, while future technologies will aim to connect different organ chips to form human-on-a-chip systems, better simulating systemic drug metabolism and toxic reactions. Such systems will integrate multiple organ models such as liver, heart, brain, and kidney, connected through microfluidic networks, providing more accurate human response predictions than traditional animal models. Technological advances in this direction will greatly change the drug development process, potentially reducing reliance on animal testing and improving drug development efficiency.

In the field of personalized medicine, cell culture technology will also play an increasingly important role. Combining patient-specific induced pluripotent stem cells (iPSCs) with advanced 3D dynamic culture systems, researchers will be able to establish patient-specific disease models for personalized drug screening and treatment plan optimization. Particularly in oncology, rapidly constructing 3D tumor models using patient tumor cells can test the effects of different drug combinations before treatment begins, providing powerful tools for precision medicine. Such applications will promote the transition of cell culture technology from basic research tools to clinical decision support means.

The role of Chinese enterprises in the future development of cell culture technology deserves special attention. Chinese biotechnology enterprises represented by Sagebio, through the successful development of the SARC series, have demonstrated an innovation model that combines local needs with global vision-1-5-9. These enterprises not only understand the specific needs and workflows of domestic researchers but can also absorb international advanced technical concepts to develop competitive products. With China's continuous investment in the biotechnology field and the continuous expansion of market demand, we have reason to believe that more Chinese enterprises will transition from technology followers to technology leaders, occupying important positions in the global cell culture equipment market.

The rise of Chinese enterprises will also promote the diversified development and accessibility improvement of global cell culture technology. By providing equivalent performance but lower-cost alternatives, Chinese enterprises enable more resource-limited laboratories to use advanced 3D dynamic culture technology, thereby promoting the progress of global scientific research. Meanwhile, specialized culture systems developed for regional diseases (such as those with high incidence in Asia) will also enrich the choice of global research tools, providing support for solving specific health problems.

The future of cell culture technology is full of promise but also faces challenges. How to better simulate the role of the immune system in tissue development and disease, how to construct vascularized tissue engineering constructs, and how to balance the complexity and usability of systems are all directions that need continued exploration. But in any case, the technological evolution path from static 2D to rotating 3D dynamic culture has pointed the way forward for us—more respect for the biological characteristics of cells, more precise simulation of the in vivo microenvironment, and more intelligent control of system operation.

8 Conclusion

The fifty-year evolution of cell culture technology is a spiral upward process from simple to complex, from static to dynamic, from two-dimensional to three-dimensional. This journey has not only witnessed the advancement of technology itself but also reflected the transformation of scientific research paradigms—from trying to simplify systems to identify basic laws to striving to build complex systems to better simulate the real world.

In the development path of cell culture technology, several key transitions are particularly evident: the transition from static monolayer culture to dynamic suspension culture improved mass transfer efficiency; the transition from two-dimensional platforms to three-dimensional scaffolds reconstructed the cellular spatial environment; the transition from homogeneous systems to heterogeneous co-culture restored cell-cell interactions; the transition from fixed parameters to adaptive control realized the optimization of the culture process. These transitions collectively point to one goal: creating a microenvironment in vitro that is as close as possible to in vivo conditions, enabling cells to exhibit their true behavior in living organisms in petri dishes.

Looking back on this history of technological evolution, we can also clearly see the role transition of China in the field of life science instruments from absentee to participant and then to innovator. From complete reliance on imported equipment in the early days, to later imitation and improvement, to today's development of products with independent intellectual property rights and international competitiveness, such as Sagebio's SARC series, this transition not only reflects the improvement of China's technological strength but also预示着 potential changes in the global life science instrument market landscape.

The advancement of cell culture technology has forever changed the way biological research is conducted. It has enabled scientists to ask and answer more complex questions, promoting the paradigm shift from molecular cell biology to systems biology. In the future, with the further maturation of technology, we expect to see cell culture play a more important role in regenerative medicine, disease modeling, and personalized medicine, ultimately achieving direct translation from laboratory research to clinical application.

Just as NASA's RWVB unexpectedly opened up new paths for ground-based cell culture technology, the future of cell culture technology may also come from breakthroughs and innovations that are difficult to foresee today. But in any case, respect for the fundamental laws of life and persistence in scientific exploration will continue to lead this field forward, making new contributions to human health and scientific progress.