Name: imaging-guided bioreactor for generating bioengineered airway tissue
Uploaded: 2022-04-06T13:50:00
Description: Watch this Scientific Journal Video about Imaging-Guided Bioreactor for Generating Bioengineered Airway Tissue at JoVE.com

这项研究得到了美国胸科学会基金会研究计划，新泽西州健康基金会和国家科学基金会（CAREER奖2143620）的部分支持。和美国国立卫生研究院（P41 EB027062）至G.V.N.

下面的动物组织方案已获得史蒂文斯理工学院动物护理和使用委员会研究所（IACUC）的动物福利指南和法规的批准，并且符合美国国立卫生研究院（NIH）关于使用实验动物的指南。
1. 影像引导大鼠气管生物反应器的设计与搭建
<ol><li>大鼠气管生物反应器的设计与制造<ol>
<li>使用 CAD 生成器软件创建具有相关设计（如入口、出口和组织培养室）的生物反应器腔室的...

<ol>
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X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+generation+of+lung+and+airway+epithelial+cells+from+human+pluripotent+stem+cells.">Efficient generation of lung and airway epithelial cells from human pluripotent stem cells.</a> Nature Biotechnology. 32 (1), 84-91 (2014).</li><li>Huang, S. X. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+in+vitro+generation+of+lung+and+airway+progenitor+cells+from+human+pluripotent+stem+cells.">The in vitro generation of lung and airway progenitor cells from human pluripotent stem cells.</a> Nature Protocols. 10 (3), 413-425 (2015).</li><li>Kim, J., O'Neill, J. D., Dorrello, N. 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在这项工作中，我们创建了一种成像引导的生物反应器，可以允许（i）在细胞去除和外源性细胞递送后 原位 监测气管腔，以及（ii）细胞接种气管组织的长期 体外 培养。使用这种定制的生物反应器，我们证明了（i）使用洗涤剂和振动辅助气道洗涤从气管腔中选择性地去除内源性上皮细胞，以及（ii）使用细胞负载的胶原I预凝胶将外源性细胞均匀分布到脱毛气管腔表面上。此外，?...

呼吸道的腔内表面衬有一层上皮，主要由多纤毛，球杆，高脚杯和基底干细胞1，2组成。上皮层是肺的主要防御机制，充当生物物理屏障，保护下面的气道组织免受吸入病原体，颗粒物或化学气体的侵害。它 通过 多种机制保护气道组织，包括细胞间紧密连接形成，粘膜纤毛清除以及抗菌和抗氧化剂分泌3，4。气道上皮缺陷与破坏性呼吸系统疾病相关，例如慢性阻塞性肺疾病 （COPD）5、原发性睫状肌运动障碍 （PCD）6 和囊性纤维化 （CF）7。
肺芯片（LOC）技术的进步代表了研究人类肺部发育，模拟各种肺部疾病以及在严格监管 的体外 环境中开发新治疗材料的机会。例如，气道上皮和内皮可以在薄的多孔膜的相对侧培养，以模拟交换肺组织的气体，从而允许忠实的疾病建模和药物测试8。同样，已经创建了 体外 疾病模型来模拟 体外气道疾病，例如COPD9 和囊性纤维化10。然而，LOC装置的一个主要挑战是重述肺组织的复杂三维（3D）结构和动态细胞 - 组织基质 相互作用在体外11。
最近，已经开发了创新的组织工程方法，允许操作 离体 肺组织12。使用这些方法， 可以通过化学， 物理和机械处理从肺组织中去除内源性细胞来制备脱落的同种异体或异种组织移植物13。此外，去细胞化肺支架中保存的天然组织细胞外基质（ECM）为植入细胞附着，增殖和分化提供了物理模拟结构，生化和生物力学线索14，15。
在这里，报道了通过结合LOC和组织工程技术创建的成像引导的生物反应器系统，以允许 体外 组织操作和外植大鼠气管组织的培养。使用这种气道组织生物反应器，该方案证明选择性地去除内源性上皮细胞，而不会破坏气道组织的潜在上皮下细胞和生化成分。接下来，我们通过滴注细胞加载的胶原I预凝胶溶液，展示新接种的外源细胞（如间充质干细胞（MSCs））在剥脱的气道腔上的均匀分布和瞬时沉积。此外，通过使用集成到生物反应器中的微光学成像装置，还可以在上皮去除和内源性细胞递送过程中实现气管腔的可视化。此外，还表明气管和新植入的细胞可以在生物反应器中培养4天而不会出现明显的细胞死亡和组织降解。我们设想，本研究中使用的成像生物反应器平台，基于薄膜的去上皮化技术以及细胞递送方法可用于生成用于 体外 疾病建模和药物筛选的气道组织。
生物反应器包括一个矩形腔室，连接到可编程注射泵，灌注泵和呼吸机，用于培养分离的大鼠气管。生物反应器具有连接到气管或组织培养室的入口和出口，以分别向气管的内部和外部空间供应试剂（例如培养基）（图1）。定制的成像系统可用于在细胞水平上可视化 体外培养的大鼠气管内部（图2）。 通过 滴注基于洗涤剂的去细胞化溶液，然后进行振动辅助气道清洗，去除气管的内源性上皮（图3）。水凝胶溶液，如I型胶原蛋白，被用作在剥脱的气管腔内均匀和瞬时接种外源细胞的递送载体（图4）。用于构建生物反应器和进行实验的所有材料均在 材料表中提供。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1&#215; PBS</td>
			<td>Gibco, Thermo Fisher Scientific</td>
			<td>10-010-031</td>
			<td></td>
		</tr>
		<tr>
			<td>3-port connector</td>
			<td>World Precision Instruments</td>
			<td>14048-20</td>
			<td></td>
		</tr>
		<tr>
			<td>4-port connector</td>
			<td>World Precision Instruments</td>
			<td>14047-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Accelerometer</td>
			<td>STMicroelectronics</td>
			<td>IIS3DWBTR</td>
			<td></td>
		</tr>
		<tr>
			<td>Achromatic doublet</td>
			<td>Thorlabs</td>
			<td>AC254-150-A-ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Aluminum pin stub</td>
			<td>TED PELLA</td>
			<td>16111</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-antimycotic</td>
			<td>Thermo Fisher Scientific</td>
			<td>15240062</td>
			<td></td>
		</tr>
		<tr>
			<td>Assembly rod</td>
			<td>Thorlabs</td>
			<td>ER1</td>
			<td></td>
		</tr>
		<tr>
			<td>Button head screws</td>
			<td>McMaster-Carr</td>
			<td>91255A274</td>
			<td></td>
		</tr>
		<tr>
			<td>Cage cube</td>
			<td>Thorlabs</td>
			<td>C4W</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbon double-sided conductive tape</td>
			<td>TED PELLA</td>
			<td>16073</td>
			<td></td>
		</tr>
		<tr>
			<td>CFSE labelling kit</td>
			<td>Abcam</td>
			<td>ab113853</td>
			<td></td>
		</tr>
		<tr>
			<td>Citrisolv (clearing agent)</td>
			<td>Decon</td>
			<td>1061</td>
			<td></td>
		</tr>
		<tr>
			<td>C-mount adapter</td>
			<td>Thorlabs</td>
			<td>SM1A9</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagen I</td>
			<td>Advanced BioMatrix</td>
			<td>5153</td>
			<td></td>
		</tr>
		<tr>
			<td>Conductive liquid silver paint</td>
			<td>TED PELLA</td>
			<td>16034</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic mirror</td>
			<td>Semrock</td>
			<td>DI03-R488</td>
			<td>Reflected laser wavelengths:&#160; 473.0 +- 2 nm 488.0 +3/-2 nm</td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified Eagle&#8217;s medium</td>
			<td>Gibco, Thermo Fisher Scientific</td>
			<td>11965118</td>
			<td></td>
		</tr>
		<tr>
			<td>Female luer bulkhead to hose barb adapter</td>
			<td>Cole-Parmer</td>
			<td>EW-45501-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Female luer to tubing barb</td>
			<td>Cole-Parmer</td>
			<td>EW-45508-03</td>
			<td></td>
		</tr>
		<tr>
			<td>Female to male luer connector</td>
			<td>Cole-Parmer</td>
			<td>ZY-45508-80</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Gibco, Thermo Fisher Scientific</td>
			<td>10082147</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter lens</td>
			<td>Chroma Technology Corp</td>
			<td>ET535/50m</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescent microscope</td>
			<td>Nikon</td>
			<td>Eclipse E1000 - D</td>
			<td></td>
		</tr>
		<tr>
			<td>Fusion 360</td>
			<td>Autodesk</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hex nut</td>
			<td>McMaster-Carr</td>
			<td>91813A160</td>
			<td></td>
		</tr>
		<tr>
			<td>Hexamethyldisilazane (HMDS)</td>
			<td>Fisher Scientifc</td>
			<td>AC120585000</td>
			<td></td>
		</tr>
		<tr>
			<td>Imaging fiber</td>
			<td>SELFOC, NSG group</td>
			<td>GRIN lens</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser</td>
			<td>Opto Engine</td>
			<td>MDL-D-488-150mW</td>
			<td></td>
		</tr>
		<tr>
			<td>Lens tubes</td>
			<td>Thorlabs</td>
			<td>SM1L40</td>
			<td></td>
		</tr>
		<tr>
			<td>LIVE/DEAD Viability/Cytotoxicity Kit (Invitrogen)</td>
			<td>Thermo Fisher Scientific</td>
			<td>L3224</td>
			<td></td>
		</tr>
		<tr>
			<td>MACH 3 CNC Control Software</td>
			<td>Newfangled Solutions</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Objective lens</td>
			<td>Olympus</td>
			<td>UCPLFLN20X</td>
			<td></td>
		</tr>
		<tr>
			<td>Peristaltic Pump</td>
			<td>Cole Parmer</td>
			<td>L/S standard digital pump system</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human FGF-basic</td>
			<td>PeproTech</td>
			<td>100-18B</td>
			<td></td>
		</tr>
		<tr>
			<td>Retaining ring</td>
			<td>Thorlabs</td>
			<td>SM1RR</td>
			<td></td>
		</tr>
		<tr>
			<td>Scientific CMOS camera</td>
			<td>PCO Panda</td>
			<td>PCO Panda 4.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulfate</td>
			<td>VWR</td>
			<td>97064-472</td>
			<td></td>
		</tr>
		<tr>
			<td>Solidworks (2019)</td>
			<td>Dassault Syst&#232;mes</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stackable lens tube</td>
			<td>Thorlabs</td>
			<td>SM1L10</td>
			<td></td>
		</tr>
		<tr>
			<td>Subwoofer plate amplifier</td>
			<td>Dayton Audio</td>
			<td>SPA250DSP</td>
			<td></td>
		</tr>
		<tr>
			<td>Subwoofer speaker</td>
			<td>Dayton Audio</td>
			<td>RSS21OHO-4</td>
			<td>Diaphragm diameter: 21 cm</td>
		</tr>
		<tr>
			<td>Syringe Pump</td>
			<td>World Precision Instruments</td>
			<td>AL-4000</td>
			<td></td>
		</tr>
		<tr>
			<td>Threaded cage plate</td>
			<td>Thorlabs</td>
			<td>CP33</td>
			<td></td>
		</tr>
		<tr>
			<td>Threaded luer adapter</td>
			<td>Cole-Parmer</td>
			<td>EW-45513-81</td>
			<td></td>
		</tr>
		<tr>
			<td>Tube lens</td>
			<td>Thorlabs</td>
			<td>AC254-150-A-ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Tygon Tubing</td>
			<td>Cole-Parmer</td>
			<td>13-200-110</td>
			<td></td>
		</tr>
		<tr>
			<td>XY Translator</td>
			<td>Thorlabs</td>
			<td>CXY1</td>
			<td></td>
		</tr>
	</tbody>
</table>

imaging-guided bioreactor for generating bioengineered airway tissue

气道组织反复损伤可损害肺功能并引起慢性肺病，如慢性阻塞性肺病。再生医学和生物反应器技术的进步为生产实验室培养的功能性组织和器官结构体提供了机会，这些结构和结构体可用于筛选药物，模拟疾病和工程组织替代物。在这里，描述了一种小型化生物反应器与成像方式相结合，该成像方式允许在体外组织操作和培养期间原位可视化外植大鼠气管的内腔。使用这种生物反应器，该方案展示了成像引导的内源性细胞成分的选择性去除，同时保留了气道组织基质的内在生化特征和超微结构。此外，还显示了外源性细胞在去细胞化气道腔上的递送，均匀分布和随后的延长培养，并进行光学原位监测。结果强调，成像引导的生物反应器可能用于促进功能性体外气道组织的产生。

基于GRIN晶状体 的原位 成像方式可以原 位 显示气管内腔（图5A）。使用这种成像方法，可以获得天然和去上皮化气管的明场和荧光图像（图5B，C）。在CFSE标记之前，没有从天然气管中观察到荧光信号（图5Bii）。然而，当用CFSE染料标记气管上皮时，在整个上皮中观察到均匀的荧光信号（绿色）（<strong cl...

Watch this Scientific Journal Video about Imaging-Guided Bioreactor for Generating Bioengineered Airway Tissue at JoVE.com

Imaging-Guided Bioreactor for Generating Bioengineered Airway Tissue

该协议描述了一种支持成像的生物反应器，该反应器允许从大鼠气管中选择性地去除内源性上皮，并在管腔表面均匀分布外源细胞，然后对细胞组织构建体进行长期 体外 培养。

成像引导的生物反应器，用于生成生物工程气道组织

Repeated injury to airway tissue can impair lung function and cause chronic lung disease, such as chronic obstructive pulmonary disease. Advances in ...

该方案允许在不同的组织操作程序（例如去上皮化和气道组织再生）期间对分离的气道组织进行体外培养和直接显微镜可视化。本研究中提出的原位成像允许在成像引导下受控切除内源性上皮和外源性细胞递送期间快速无损地监测气管腔。成像引导的生物反应平台和组织操作方案可用于生成生物工程气道组织，用于疾病建模和药物筛选。成像气道组织生物反应器的构建和使用将由我们研究小组的两名博士生Mohammad Mir和Jiawen Chen演示。要开始创建原位成像设备，请将镜筒透镜插入可堆叠透镜管中，并使用挡圈将其固定。然后，通过C接口适配器将镜头台组件安装到科学CMOS相机上。使用Micromanager软件操作相机并获取照片和视频。在瞄准距离相机10米的物体时，调整管透镜与摄像机成像传感器之间的距离，直到在计算机屏幕上形成物体的聚焦图像。接下来，使用光学笼系统组件，如装配杆，螺纹笼板和笼式立方体，将滤光透镜安装在双刃，超分辨率，二向色镜和激光器上。将20倍物镜连接到设备。然后，通过X-Y转换器在镜管的远端安装直径为500微米的绿色透镜。调整绿色和物镜之间的距离，形成聚焦的微观图像。为了在明场或荧光中可视化分离的大鼠气管腔，将生物反应器放在成像台上。接下来，注入500微升新鲜制备的羧基荧光素琥珀酰亚胺酯或CFSE溶液，流速为每分钟5毫升，通过连接到气管套管的注射泵通过气管。一旦 CFSE 溶液充满气管，停止泵。10分钟后，使用注射泵注入10毫升PBS以除去残留的未掺入的CFSE试剂，以洗涤气管腔。洗涤后，通过连接到气管一端的鲁尔连接器将绿色晶状体的远端成像端插入气管。然后，轻轻地将绿色晶状体移入气管内，直到气管表面聚焦。要在显微管理器软件中捕获明场图像，请通过笼状立方体用白光照亮气管腔。然后，单击“实时”图标以实时显示气管的管腔表面。使用“成像”设置和“曝光”选项卡将曝光时间更改为所需值。要调整图像的对比度和亮度，请使用直方图和强度缩放窗口在交互式直方图显示的端点处移动黑白箭头。单击“停止实时”以激活“捕捉”图标，然后单击“捕捉”图标以冻结图像。接下来，使用“导出”和“图像作为置换选项卡”设置，以所需格式保存图像。要在荧光模式下获取照片和视频，请通过笼状立方体用CFSE专用激光照亮气管腔，并通过来回移动成像探头实时获取图像。获得照片和视频后，轻轻地从气管中取出绿色晶状体。对于气管的去上皮化，通过气管套管滴入50微升新鲜制备的2%十二烷基硫酸钠，以在气管腔上产生洗涤剂溶液的薄膜。滴注后，使用公头或母头鲁尔塞关闭生物反应器的管道连接，并将生物反应器转移到37摄氏度的培养箱中10分钟，以使SDS停留在气管内。重复滴注一次。第二次滴注后，通过注射泵以每分钟10毫升的流速用500微升PBS冲洗气管腔三次，以去除裂解的上皮和SDS。然后，将生物反应器放在摇床上，以20赫兹的频率和0.3毫米的位移幅度进行机械振动，以物理方式促进SDS处理的上皮细胞从气管腔中分离。当气管被机械振动时，通过气管腔滴注500微升PBS两次，以除去残留的SDS和细胞碎片。去除上皮后，通过使用绿色晶状体成像装置测量CFSE的强度来评估上皮层的清除率。制备去上皮化大鼠气管后，在37摄氏度的水浴中解冻冷冻的间充质干细胞或MSCs30秒。然后，用血细胞计数器计数细胞，然后制备浓度为每毫升6个细胞的5倍10倍的细胞溶液。然后，通过在室温下用两毫升100微摩尔CFSE溶液孵育细胞来荧光标记细胞。15分钟后，用五毫升PBS冲洗细胞三次，然后将其重悬于新鲜的DMEM培养基中，最终浓度为每毫升10倍至7个细胞。如手稿所述，在制备胶原I水凝胶后，立即将细胞加入具有所需浓度的水凝胶溶液中。然后，将细胞和凝胶溶液与微量移液管混合，得到均匀的细胞 - 水凝胶混合物。接下来，通过鲁尔连接器将生物反应器内气管的一端连接到可编程注射泵上，并将五毫升新鲜培养基以37摄氏度的速度输送到生物反应器室中，以覆盖气管的外表面。然后，将10微升的细胞 - 水凝胶混合物推注到生物反应器内的去上皮化气管中，以在气管腔上产生细胞水凝胶层。细胞注射后，将生物反应器置于37摄氏度和5%二氧化碳的无菌细胞培养箱中进行烧制30分钟以进行糖化，以进行糖化。为了可视化植入细胞的分布，用70%异丙醇或乙醇对绿色晶状体进行灭菌，并将生物反应器放在成像台上，以获得明场和荧光模式下的照片和视频。细胞接种30分钟后，以每分钟一毫升的流速将一毫升培养基注入气管腔中。最后，在37摄氏度的培养箱中在生物反应器内培养细胞接种的气管，以达到所需的时间。在细胞培养过程中，保持管腔内的培养基不变，而气管外的培养基通过单向流动连续灌注。在去上皮化气管的明场和荧光图像中，在CFSE标记之前没有观察到荧光信号。使用CFSE，在整个上皮细胞中观察到均匀的荧光信号。去上皮化后，荧光强度显着降低，表明上皮消融。去上皮化气管的苏木精和曙红染色表明，从气管腔中去除假分层上皮，并在下层组织层中保存细胞和细胞外基质或ECM微结构。此外，五色和三色染色证实了气管组织结构和ECM成分（如胶原蛋白和蛋白聚糖）的维持。上皮细胞和胶原I的免疫荧光显示上皮完全去除并保留上皮下组织内的胶原I。扫描电子显微照片显示，天然气管中充满了多毛细胞和高脚杯细胞。在去上皮化的气管中，基底膜暴露出来，如细胞外基质纤维的网状网络和上皮细胞的缺失所示。去除上皮化气管的荧光图像，在接种PBS和胶原蛋白时，如下所示。与通过培养基接种的细胞相比，通过水凝胶递送的荧光标记细胞在管腔内保持更均匀的粘附。细胞活力研究表明，细胞递送程序对细胞活力没有影响，超过90%的细胞仍然存活。在去上皮化过程中创建洗涤剂薄膜并使用水凝胶作为细胞的递送载体是该方案成功的重要步骤。我们的下一个研究目标是将实验室培养的气道原代或干细胞植入去上皮化的气道组织上，以研究是否可以制备功能性气道组织。

Research

JoVE Journal

Bioengineering

Design of a Biaxial Mechanical Loading Bioreactor for Tissue Engineering

组织工程生物反应器的双轴机械装载设计

We  designed a loading device that is capable of applying uniaxial or biaxial  mechanical strain to a tissue engineered biocomposites fabricated for  ...

科研

生物工程

Microfluidic Picoliter Bioreactor for Microbial Single-cell Analysis: Fabrication, System Setup, and Operation

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described ...

Operation of a Benchtop Bioreactor

一个台式生物反应器的操作

Fermentation systems are used to provide an optimal growth environment for many different types of cell cultures. The ability afforded by fermentors to ...

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

对于中吸入有毒化学测试体外细胞培养模型

Cell cultures are indispensable to develop and study efficacy of therapeutic agents, prior to their use in animal models. We have the unique ability to ...

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

适应静电过程中提供三个独特的环境为一个三层次<em&gt;在体外</em&gt;的气道壁模型

Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to ...

Generation of ESC-derived Mouse Airway Epithelial Cells Using Decellularized Lung Scaffolds

用脱细胞支架龙ESC衍生鼠气道上皮细胞的生成

Lung lineage differentiation requires integration of complex environmental cues that include growth factor signaling, cell-cell interactions and ...

3D Magnetic Stem Cell Aggregation and Bioreactor Maturation for Cartilage Regeneration

3D磁干细胞聚集和生物反应器为成熟软骨再生

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach ...

Recombinant Collagen I Peptide Microcarriers for Cell Expansion and Their Potential Use As Cell Delivery System in a Bioreactor Model

重组胶原蛋白 i. 肽载体在生物反应器中的应用及其作为细胞传递系统的潜力

Tissue engineering is a promising field, focused on developing solutions for the increasing demand on tissues and organs regarding transplantation ...

Cardiac Spheroids as in vitro Bioengineered Heart Tissues to Study Human Heart Pathophysiology

心脏球形作为体外生物工程心脏组织研究人类心脏病理生理学

Despite several advances in cardiac tissue engineering, one of the major challenges to overcome remains the generation of a fully functional vascular ...

Multi-Stream Perfusion Bioreactor Integrated with Outlet Fractionation for Dynamic Cell Culture

多流灌注生物反应器集成了出口分馏功能，用于动态细胞培养

Certain cell and tissue functions operate within the dynamic time scale of minutes to hours that are poorly resolved by conventional culture systems. This ...