Name: scalable generation of mature cerebellar organoids from human pluripotent stem cells and characterization by immunostaining
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Description: Watch this Scientific Journal Video about Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining at JoVE.com

这项工作得到了葡萄牙国家电信协会（FCT）的支持（UIDB/04565/2020通过Lisboa区域项目2020，项目N.007317， PD/BD/105773/2014 至 T.P.S 和 PD/BD/128376/2017 到 D.E.S.N.），项目由 FEDER （POR Lisboa 2020 – 项目区域行动区域葡萄牙 2 和 FCT 通过授予 PAC - 精确 LISBOA - 01 - 0145 - Feder - 016394 和 CEREBEX 生成小脑有机体为 Ataxia 研究授予 LISBOA -01-0145-FEDER-029298。还根据赠款协议第739572号——再生和精密医学发现中心H2020-2020-01-2016-2017年，从欧洲联盟的Horizon 2020研究和创新方案获得资金。

1. 在单层文化中传传和维护人类 iPSC
<ol><li>板材的制备<ol>
<li>在4°C下 解冻基底膜基质（见材料表）库存，制备60μL等分。将等分在-20°C下冷冻。</li>
		<li>要覆盖6孔板的孔，在冰上解冻地下室膜基质的一个等分。解冻后，将 DMEM-F12 的 60 μL 添加到 6 mL。通过上下移液轻轻重新暂停。</li>
		<li>将 1 mL 稀释的地下膜基质溶液添加到 6 孔板的每个孔中，并在 RT 中孵育至少 1 小时，然后以 4°C...

<ol>
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需要大量的细胞数量以及定义的培养条件来生成特定的细胞类型，用于药物筛选和再生医学应用，这一直在推动可扩展培养系统的发展。近年来，几组报告了可扩展的神经祖代和功能神经元32、33、34等34，为神经退行性疾病新模型的发展提供了重大进展。32,然而，胚胎发育的一些关键事件的回顾仍然缺乏，长期维持在悬浮过程?...

人类多能干细胞（PSCs）的出现是再生医学和疾病建模的极佳工具，因为这些细胞可以分化成人体1、2的多数细胞系。自从他们发现以来，PSC分化使用不同的方法被报告为不同的疾病建模，包括神经退行性疾病3，4，5，6。,4,5,6
最近，有报道说，3D培养物来自类似于人类大脑结构的PSC;这些被称为大脑器官3，7，8。,83,这些结构的生成来自健康和患者特定的PSC提供了一个宝贵的机会来模拟人类发展和神经发育障碍。然而，用于生成这些组织良好的脑结构的方法很难应用于其大规模生产。为了产生足够大的结构，以重述组织形态，而不在器官内坏死，协议依赖于在静态条件下的初始神经承诺，然后封装在水凝胶和随后的培养在动态系统3。然而，这种办法可能限制器官生产的潜在扩大。尽管已经努力将PSC分化到中枢神经系统的特定区域，包括皮质、神经、中脑,和脊髓神经元9、10、11、12，,10但动态条件下的特定大脑区域的生成仍然是一个挑战。11,12特别是，3D结构中成熟的小脑神经元的生成尚未描述。Muguruma等人开创了培养条件的产生，重新概括了早期小脑发育13，最近报告了一个方案，允许人类胚胎干细胞产生一个极化结构，让人联想到头三个月小脑7。然而，在报告研究中，小脑神经元的成熟需要器官分离，小脑祖细胞的排序，并在,单层培养系统7、14、15、16,14中与馈养细胞15共同培养。因此，在定义的条件下，用于疾病建模所需的小脑器官的可重复生成仍然是与培养和馈送源变异相关的挑战。
该协议提供了使用一用垂直轮生物反应器（参见规格材料表），即使用一种使用垂直轮生物反应器将人类PSC有效分化成小脑神经元Table of Materials的最佳培养条件，以下简称生物反应器。生物反应器配有大型垂直叶轮，与U形底部结合，在容器内提供更均匀的剪切分布，允许温和、均匀的混合和颗粒悬浮，降低搅拌速度17。有了这个系统，可以获得形状和大小控制的细胞聚合体，这对更均匀、更高效的分化非常重要。此外，更多的 iPSC 衍生器官可以以不太费力的方式生成。
器官的主要特征是通常由干细胞形成的3D多细胞结构，是不同细胞类型的自组织，形成特定形状，如人类形态18、19、20,19,中所看到的形状。因此，有机形态是分化过程中需要评价的重要标准。使用一组特定抗体对器官进行冷冻和进一步免疫渗透，使分子标记在空间上可视化，以分析细胞增殖、分化、细胞种群特性和凋亡。有了这个协议，通过免疫控制器官冷冻，在分化的第7天观察到了一个初始有效的神经承诺。在分化过程中，观察到几个具有小脑特征的细胞群。在这个动态系统中35天后，小脑神经皮皮沿着一个轴线组织，有一个增殖的祖细胞和基本定位的后层神经元。在成熟过程中，从第35~90天的分化，可以看到不同类型的小脑神经元，包括Purkinje细胞（卡尔宾丁+），颗粒细胞（PAX6+/MAP2+），高尔基细胞（神经粒蛋白+），单极刷细胞（TBR2+），和深小脑核投影+神经元（TBR1+）。+此外，在培养的90天后，在生成的小脑器官中观察到不重要的细胞死亡量。
在这个系统中，人类 iPSC 衍生的器官成熟成不同的小脑神经元，存活长达 3 个月，无需分离和喂食共培养，为疾病建模提供了人类小脑神经元的来源。

<table>
	<tbody>
		<tr>
			<td>3MM paper</td>
			<td>WHA3030861</td>
			<td>Merck</td>
			<td></td>
		</tr>
		<tr>
			<td>Accutase</td>
			<td>A6964 - 500mL</td>
			<td>Sigma</td>
			<td>cell detachment medium</td>
		</tr>
		<tr>
			<td>Anti-BARHL1 Antibody</td>
			<td>HPA004809</td>
			<td>Atlas Antibodies</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Calbindin D-28k Antibody</td>
			<td>CB28</td>
			<td>Millipore</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-MAP2 Antibody</td>
			<td>M4403</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-N-Cadherin Antibody</td>
			<td>610921</td>
			<td>BD Transduction</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-NESTIN Antibody</td>
			<td>MAB1259-SP</td>
			<td>R&#38;D</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-OLIG2 Antibody</td>
			<td>MABN50</td>
			<td>Millipore</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-PAX6 Antibody</td>
			<td>PRB-278P</td>
			<td>Covance</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-SOX2 Antibody</td>
			<td>MAB2018</td>
			<td>R&#38;D</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-TBR1 Antibody</td>
			<td>AB2261</td>
			<td>Millipore</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-TBR2 Antibody</td>
			<td>ab183991</td>
			<td>Abcam</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-TUJ1 Antibody</td>
			<td>801213</td>
			<td>Biolegend</td>
			<td></td>
		</tr>
		<tr>
			<td>Apo-transferrin</td>
			<td>T1147</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>BrainPhys Neuronal Medium N2-A &#38; SM1 Kit</td>
			<td>5793 - 500mL</td>
			<td>Stem cell tecnhnologies</td>
			<td></td>
		</tr>
		<tr>
			<td>Chemically defined lipid concentrate</td>
			<td>11905031</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslips 24x60mm</td>
			<td>631-1575</td>
			<td>VWR</td>
			<td></td>
		</tr>
		<tr>
			<td>Crystallization-purified BSA</td>
			<td>5470</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>10236276001</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Dibutyryl cAMP</td>
			<td>SC- 201567B -500mg</td>
			<td>Frilabo</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM-F12</td>
			<td>32500-035</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>A3840001</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin from bovine skin</td>
			<td>G9391</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Copling Jar</td>
			<td>E94</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Glutamax I</td>
			<td>10566-016</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>MB014001</td>
			<td>NZYtech</td>
			<td></td>
		</tr>
		<tr>
			<td>Ham&#8217;s F12</td>
			<td>21765029</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Human Episomal iPSC Line</td>
			<td>A18945</td>
			<td>ThermoFisher</td>
			<td>iPSC6.2</td>
		</tr>
		<tr>
			<td>IMDM</td>
			<td>12440046</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin</td>
			<td>91077C</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>iPS DF6-9-9T.B</td>
			<td></td>
			<td>WiCell</td>
			<td></td>
		</tr>
		<tr>
			<td>Iso-pentane</td>
			<td>PHR1661-2ML</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Ascorbic acid</td>
			<td>A-92902</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Matrigel</td>
			<td>354230</td>
			<td>Corning</td>
			<td>basement membrane matrix</td>
		</tr>
		<tr>
			<td>Monothioglycerol</td>
			<td>M6154</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Mowiol</td>
			<td>475904</td>
			<td>Millipore</td>
			<td>mounting medium</td>
		</tr>
		<tr>
			<td>mTeSR1</td>
			<td>85850 -500ml</td>
			<td>Stem cell technologies</td>
			<td></td>
		</tr>
		<tr>
			<td>N2 supplement</td>
			<td>17502048</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Neurobasal</td>
			<td>12348017</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>158127</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS-0.1 Single-Use Vessel</td>
			<td>SKU: IA-0.1-D-001</td>
			<td>PBS Biotech</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS-MINI MagDrive Base Unit</td>
			<td>SKU: IA-UNI-B-501</td>
			<td>PBS Biotech</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human BDNF</td>
			<td>450-02</td>
			<td>Peprotech</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human bFGF/FGF2</td>
			<td>100-18B</td>
			<td>Peprotech</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human FGF19</td>
			<td>100-32</td>
			<td>Peprotech</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human GDNF</td>
			<td>450-10</td>
			<td>Peprotech</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant human SDF1</td>
			<td>300-28A</td>
			<td>Peprotech</td>
			<td></td>
		</tr>
		<tr>
			<td>ROCK inhibitor Y-27632</td>
			<td>72302</td>
			<td>Stem cell technologies</td>
			<td></td>
		</tr>
		<tr>
			<td>SB431542</td>
			<td>S4317</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>S7903</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>SuperFrost Microscope slides</td>
			<td>12372098</td>
			<td>ThermoFisher</td>
			<td>adhesion microscope slides</td>
		</tr>
		<tr>
			<td>Tissue-Tek O.C.T. Compound</td>
			<td>25608-930</td>
			<td>VWR</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-HCL 1M</td>
			<td>T3038-1L</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>9002-93-1</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween-20</td>
			<td>P1379</td>
			<td>Sigma</td>
			<td></td>
		</tr>
		<tr>
			<td>UltraPure 0.5M EDTA, pH 8.0</td>
			<td>15575020</td>
			<td>ThermoFisher</td>
			<td></td>
		</tr>
	</tbody>
</table>

scalable generation of mature cerebellar organoids from human pluripotent stem cells and characterization by immunostaining

小脑在维持平衡和运动协调中起着至关重要的作用，不同小脑神经元的功能缺陷可以触发小脑功能障碍。目前关于疾病相关神经元表型的知识大部分基于死后组织，这使得对疾病进展和发育的理解变得困难。动物模型和不朽的细胞系也被用作神经退行性疾病的模型。然而，他们不能完全概括人类疾病。人类诱导多能干细胞（iPSCs）在疾病建模方面潜力巨大，为再生方法提供了宝贵的来源。近年来，来自患者衍生的 iPSC 的脑器官的生成改善了神经退行性疾病建模的前景。然而，在三D培养系统中，缺乏产生大量器官和高产成熟神经元的协议。所提出的协议是使用可扩展的一次使用生物反应器在化学定义条件下可重复和可扩展生成人类 iPSC 衍生器官的新方法，其中有机体获得小脑特性。生成的器官的特点是在mRNA和蛋白质水平上表达特定的标记。对特定蛋白质组的分析可以检测不同的小脑细胞群，其定位对于组织结构的评价非常重要。器官冷冻和进一步免疫污染器官切片用于评估特定小脑细胞群的存在及其空间组织。

该协议是通过使用0.1 L生物反应器促进细胞聚合（图1A）启动的。对 iPSC 进行单细胞接种，在 60 mL 的培养中播种 250，000 个细胞/mL，搅拌速度为 27 rpm。这被定义为第 0 天。24小时后，细胞有效形成球形状聚集体（第1天，图1B），形态学一直保持良好，直到第5天，大小逐渐增加，表明随着时间的推移，在聚合形态和大小方面具有高度的同质性。（<strong cla...

Watch this Scientific Journal Video about Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining at JoVE.com

Scalable Generation of Mature Cerebellar Organoids from Human Pluripotent Stem Cells and Characterization by Immunostaining

该协议描述了一个动态培养系统，用于利用一种一种用途的生物反应器，产生控制大小的人类多能干细胞，进一步刺激在化学定义和免饲料条件下小脑器官的分化。

人类多能干细胞和免疫损伤的成熟小脑有机体可伸缩生成

The cerebellum plays a critical role in the maintenance of balance and motor coordination, and a functional defect in different cerebellar neurons can ...

我们首次提出了一种可重复和可扩展的生成新方法，即在小脑有机体中衍生的 IPS 细胞，以及使用一次使用生物反应器的化学定义条件。为了获得用于播种生物反应器的细胞，在人类 iPSC 六井板的每个井中加入一毫升细胞分离介质。在37摄氏度下孵育板7分钟，直到轻轻的震动很容易将细胞从井中分离。使用P 1000微移液器，上下移液细胞分离介质，直到细胞分离成单个细胞。接下来添加两毫升，完整的细胞培养培养培养能力到每个井，这将灭活酶消化。然后使用移液器，轻轻地将细胞转移到无菌锥形管中。在重力210倍时将管离心三分钟。取出上一提液，并在培养培养中重新悬浮细胞颗粒。使用血细胞计计算细胞数，并尝试蓝色染料。见生物反应器容器与 iPSC 的，密度为每毫升25万细胞。将生物反应器容器插入通用基座单元，放入37摄氏度、95%湿度和5%二氧化碳的孵化器中。要促进 iPSC 聚合，请将基本控制单元的搅拌速率设置为 27 RPM。第一天，第1天是播种生物反应器的日子，将生物反应器和基础单元放在无菌流罩中，然后使用血清移液器，收集一毫升样品，细胞悬浮液。将电池悬浮板放在超低附件中，24个井板。使用显微镜，确认 iPSC 衍生的聚合已经形成。如果是这样，以40 X或100 X捕捉集料的图像。当集料，达到200至250微米的直径，让所有的器官沉淀，在生物反应器的底部，然后更换所有花费的介质，与g方体分型介质。将基座和生物反应器放在培养箱中，将搅拌降低至 25 RPM。从储存的有机物中去除上经剂后，在颗粒中加入1毫升15%蔗糖，然后轻轻旋转，混合均好。在四摄氏度下孵育有机体后，取出蔗糖溶液，然后加入15%蔗糖、7.5%明胶的一毫升，然后轻轻旋转快速混合。当有机体在37摄氏度下孵育时，用15%蔗糖7.5%明胶溶液填充一个塑料容器，并允许其凝固。在有机体孵育一小时后，使用巴斯德移液器，将一滴有机糖明胶混合物放在凝固蔗糖和明胶上。等待15分钟后，下降凝固，填补塑料容器的其余部分，用更多的15%蔗糖，7.5%明胶，允许它完全凝固在室温下，然后在4摄氏度下孵育20分钟。将明胶切成立方体，中间有有机物，将立方体固定成一块纸板，用一滴 OCT 化合物，然后将 250 毫升异丙烷放入 500 毫升的杯子中，用液氮填充适当的容器。使用钳子和厚手套，小心地将异丙烷杯放在液氮表面。异丙烷和液氮是危险试剂。使用这两种试剂时，需要个人防护设备，包括厚手套和充足的通风。当异素烷冷却到负80摄氏度时，将明胶立方体放在异素烷中，直到它冻结。一个冰冻良好的立方体，在用钳子轻轻敲击时产生金属声音，表明它已准备好存储。用 50 毫升 1X PBS 清洗包含器官部分的显微镜幻灯片，5 分钟，然后将幻灯片转移到包含新鲜 1X PBS 的科普林罐中。将幻灯片转移到含有 50 毫升新鲜准备的甘氨酸的科普林罐中，在室温下孵育 10 分钟，然后将幻灯片转移到含有 0.1%Triton 的 50 毫升的科普林罐中，并在室温下渗透 10 分钟。用 1X PBS 清洗幻灯片两次，每次洗涤 5 分钟。准备免疫浸盘子，用三毫米纸浸泡在1X PBS中。用切片周围的纸巾擦干幻灯片，并把它们放在三毫米纸上。使用巴斯德移液器，用 0.5 毫升的阻尼溶液盖住每张幻灯片。在室温下孵育30分钟后，去除多余的阻塞溶液，用纸巾驱动幻灯片，在切片周围。将稀释原抗体的50微升放在每张幻灯片上，用盖板盖住它们。将滑梯放在先前准备的免疫抹盘中，并在摄氏四度时孵育。经过一夜的孵育，将幻灯片转移到一个Coplin罐子，50毫升的TBST，让盖板脱落，然后用TBST洗三次幻灯片，每次5分钟。将稀释的二次抗体的50微升放在每张幻灯片上，并盖上盖板。将幻灯片放在先前准备的免疫抹盘中，在室温下孵育30分钟，防止光线。将幻灯片再次转移到科普林罐中，用 50 毫升 TBST 洗涤三次，每次洗涤五分钟。使用巴斯德移液器，在0.5毫升的dappy溶液，在每个幻灯片的整个表面，并在室温下孵育5分钟。在生物反应器中24小时后，iPSC有效地形成了球形状的集料。形态在第五天之前一直保持良好，尺寸逐渐扩大。表现出高度的同质性。显微镜的定量分析还显示，第一天聚合大小的正态分布，两个细胞系在第二天达到最佳的聚合大小。在达到所需的聚合直径后，诱导神经承诺，然后生成不同的小脑祖细胞。在分化器官过程中，显示一种更明显的上皮化，类似于具有发光空间的神经管状结构。此外，器官直径分布，在初始小脑承诺期间均匀，直到第14天。免疫荧光分析支持，一个有效的神经承诺，iPSC衍生的器官，已经实现，第七天分化。免疫污染也证明，有效的小脑承诺和小脑器官的高效成熟。此外，在生物反应器80天后，有机物的活死染色显示出高细胞活性，并且没有证据表明坏死区。这项技术是研究病理通路的重要工具，它涉及小脑退化，并评估新药的治疗效果。

Research

JoVE Journal

Bioengineering

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and Induced Pluripotent Stem Cells (iPSCs)

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells hESCs and Induced Pluripotent Stem Cells iPSCs

开发，扩展，<em&gt;在体内</em&gt;从人类胚胎干细胞（hESCs）监测人类NK细胞和诱导多能干细胞（iPS细胞）

We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to ...

科研

生物工程

Transplantation of Induced Pluripotent Stem Cell-derived Mesoangioblast-like Myogenic Progenitors in Mouse Models of Muscle Regeneration

在肌肉再生的老鼠模型中移植诱导多能干细胞衍生的中生代细胞样肌原体

Patient-derived iPSCs could be an invaluable source of cells for future autologous cell therapy protocols. iPSC-derived myogenic stem/progenitor cells ...

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System

利用 CRISPR-Cas9 系统高效生成和编辑人胰细胞无饲养干细胞

Embryonic and induced pluripotent stem cells can self-renew and differentiate into multiple cell types of the body. The pluripotent cells are thus coveted ...

Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate (LAL) Assays

Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate LAL Assays

用利穆鲁布莱巴细胞裂解液 (lal) 法检测纳米配方中的内毒素

When present in pharmaceutical products, a Gram-negative bacterial cell wall component endotoxin (often also called lipopolysaccharide) can cause ...

Growth and Characterization of Irradiated Organoids from Mammary Glands

乳腺辐照有机体的生长与表征

Organoids derived from the digested tissue are multicellular three-dimensional (3D) constructs that better recapitulate in vivo conditions than cell ...

Isolation of Adult Human Dermal Fibroblasts from Abdominal Skin and Generation of Induced Pluripotent Stem Cells Using a Non-Integrating Method

使用非整合方法分离成人人体皮肤纤维细胞与诱导多能干细胞的生成

Induced pluripotent stem cells (iPSCs) could be considered, to date, a promising source of pluripotent cells for the management of currently untreatable ...

Guided Differentiation of Mature Kidney Podocytes from Human Induced Pluripotent Stem Cells Under Chemically Defined Conditions

在化学定义条件下，成熟肾细胞与人类诱导多能干细胞的引导分化

Kidney disease affects more than 10% of the global population and costs billions of dollars in federal expenditures. The most severe forms of kidney ...

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

人类诱导多能干细胞衍生心肌细胞中的单细胞光学作用潜在测量

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically ...

Fibroblast Derived Human Engineered Connective Tissue for Screening Applications

成纤维细胞衍生的人类工程结缔组织用于筛查应用

Fibroblasts are phenotypically highly dynamic cells, which quickly transdifferentiate into myofibroblasts in response to biochemical and biomechanical ...