Name: efficient and scalable directed differentiation of clinically compatible corneal limbal epithelial stem cells from human pluripotent stem cells
Uploaded: 2018-10-24T14:00:00
Description: Watch this Scientific Journal Video about Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells at JoVE.com

该研究得到了芬兰科学院 (赠款297886号)、特克斯的人力备件计划、芬兰技术和创新资助机构、芬兰眼和组织银行基金会以及芬兰文化基金会的支持。作者感谢生物医学实验室技术人员我 Melin、汉娜 Pekkanen、艾玛维克斯泰特和我 Heikkilä为卓越的技术援助和对细胞培养的贡献。Katriina 教授 Setälä被公认为提供用于荧光成像设备的 hiPSC 线和普司通成像核心设施。

坦佩雷大学经芬兰国家法医学鉴定事务局 (Dnro 1426/32/300/05) 批准, 对人类胚胎进行研究。该研究所还有皮尔卡曼医院区道德委员会的支持性声明, 以推导、培养和区分人类胚胎干细胞线 (Skottman/R05116), 并在眼科研究中使用 hiPSC 线 (Skottman/R14023)。这项研究没有新的细胞系。
注意: 所述的协议是基于特定的, 商业上可用的 hPSC 和角膜上皮分化培养基。有关制造商/供应商信息和目录编...

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该协议的预期结果是在大约3.5 周内从 FF hPSC 的单个电池悬浮液中成功和稳健地生成 LESCs。由于角膜上皮发育从表面外胚层29, 该议定书的第一步旨在指导 hPSCs 走向这一血统。24 h 诱导与转化生长因子β (SB-505124) 拮抗剂, 和 bFGF 用于诱导外胚层分化, 其次 48 h 中胚层 BMP-4 提示推动细胞朝向表面外胚层。将 IV/LN-521 组合矩阵的以下贴壁分化步骤与化学定义的分化培养基结合在一起, 进?...

眼睛表面的透明角膜允许光线进入视网膜并提供大部分眼球的折射力。角膜缘上皮干细胞 (LESCs) 持续再生, 是最外层的层状角膜上皮细胞。LESCs 位于巩膜交界处1,2的角膜缘壁龛的基底层。LESCs 缺乏特定的和独特的标记, 因此它们的识别需要对一套假定标记进行更广泛的分析。上皮转录因子 p63, 特别是 p63 (ΔNp63α) 阿尔法异构体的 n-末期截断转录, 已被提出作为一个相关的阳性 LESC 标记3,4。LESCs 的不对称分裂允许他们自我更新, 但也产生迁移向心地和前方的后代。随着细胞向角膜表面的进展, 他们逐渐失去了干性, 最后最终分化为从角膜表面不断丢失的表面鳞状细胞。
对任何角膜层的损伤都会导致严重的视力损害, 因此角膜缺陷是全球视力丧失的主要原因之一。在角膜缘干细胞缺乏症 (康乐文化署), 角膜缘被破坏的疾病或创伤导致 conjunctivalization 和白内障的表面和随后的视力损失5,6。使用自体或异基因角膜缘移植的细胞置换疗法为康乐文化署4、7、8、9的患者提供治疗策略。然而, 收获自体移植有并发症的风险, 健康的眼睛, 捐助组织是供不应求。人类多能干细胞 (hPSCs), 特别是人类胚胎干细胞 (hESCs) 和人类诱导的多能干细胞 (hiPSCs), 可以作为一种无限的临床相关细胞类型, 包括角膜上皮细胞的来源。因此, hPSC 衍生的 LESCs (hPSC LESCs) 代表了一种吸引眼球细胞替代疗法的新细胞源。
传统上, 未分化的 hPSC 培养方法和 LESCs 的分化协议依赖于未定义的馈线细胞、动物血清、条件培养基或羊膜10、11、12,13,14,15. 最近, 对更安全的细胞治疗产品的努力促使人们寻求更标准化和异种的文化和差异化协议。因此, 对未分化 hPSCs 的长期培养的几种定义和异种的方法现在在商业上可用16,17,18。作为一个连续的, 定向分化协议依赖分子线索引导 hPSCs 角膜上皮的命运最近介绍了19,20,21,22, 23。然而, 许多这些协议使用的是未定义的, 以馈线为基础的 hPSCs 作为起始材料, 或复杂, 异种生长因子鸡尾酒的分化。
本协议的目的是提供一个强大的, 优化的, 异种和无饲养的 hPSC 培养方法和随后分化的角膜 LESCs。多能 hPSCs 在 laminin-521 (LN-521) 基质中的单层培养在定义的无白蛋白 hPSC 培养基 (特别重要的 8 Flex) 允许快速生产均匀起始材料的分化。此后, 一个简单的, 两步分化策略引导 hPSCs 朝向表面外胚层的命运在悬浮, 其次是贴壁分化到 LESCs。在24天内获得 > 65% express ΔNp63α的细胞种群。异种和无馈线协议已通过几个 hPSC 线 (hESCs 和 hiPSCs) 进行了测试, 没有任何对细胞系特定优化的要求。此处介绍的无周末维护、传代、cryostoring 和 hPSC LESC 分型协议可为临床或研究目的生产大量高质量 LESCs。

<table>
	<tbody>
		<tr>
			<td>Material/Reagent</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1x DPBS containing Ca2+ and Mg2+</td>
			<td>Gibco</td>
			<td>#14040-091</td>
			<td></td>
		</tr>
		<tr>
			<td>1x DPBS without Ca2+ and Mg2+</td>
			<td>Lonza</td>
			<td>#17-512F/12</td>
			<td></td>
		</tr>
		<tr>
			<td>100 mm cell culture dish</td>
			<td>Corning CellBIND</td>
			<td>#3296</td>
			<td>Culture vessel format for adherent hPSC-LESC differentiation</td>
		</tr>
		<tr>
			<td>12-well plate</td>
			<td>Corning CellBIND</td>
			<td>#3336</td>
			<td>Culture vessel format for IF samples</td>
		</tr>
		<tr>
			<td>24-well plate</td>
			<td>Corning CellBIND</td>
			<td>#3337</td>
			<td>Culture vessel format for IF samples</td>
		</tr>
		<tr>
			<td>2-mercaptoethanol</td>
			<td>Gibco</td>
			<td>#31350-010</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well plate, Ultra-Low attachment</td>
			<td>Corning Costar</td>
			<td>#3471</td>
			<td>Culture vessel format for induction in suspension culture</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 anti-mouse Ig</td>
			<td>ThermoFisher Scientific</td>
			<td>#A-21202</td>
			<td>Secondary antibody for IF</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 anti-rabbit Ig</td>
			<td>ThermoFisher Scientific</td>
			<td>#A-21206</td>
			<td>Secondary antibody for IF</td>
		</tr>
		<tr>
			<td>Alexa Fluor 568 anti-goat Ig</td>
			<td>ThermoFisher Scientific</td>
			<td>#A-11057</td>
			<td>Secondary antibody for IF</td>
		</tr>
		<tr>
			<td>Alexa Fluor 568 anti-mouse Ig</td>
			<td>ThermoFisher Scientific</td>
			<td>#A-10037</td>
			<td>Secondary antibody for IF</td>
		</tr>
		<tr>
			<td>Basic fibroblast growth factor (bFGF, human)</td>
			<td>PeproTech Inc.</td>
			<td>#AF-100-18B</td>
			<td>Animal-Free Recombinant Human FGF-basic (154 a.a.)</td>
		</tr>
		<tr>
			<td>BD Cytofix/Cytoperm Fixation/Permeabilization Solution</td>
			<td>BD Biosciences</td>
			<td>#554722</td>
			<td>Fixation and permeabilization solution for flow cytometry</td>
		</tr>
		<tr>
			<td>BD Perm/Wash Buffer</td>
			<td>BD Biosciences</td>
			<td>#554723</td>
			<td>Washing buffer for flow cytometry</td>
		</tr>
		<tr>
			<td>Blebbistatin</td>
			<td>Sigma-Aldrich</td>
			<td>#B0560</td>
			<td></td>
		</tr>
		<tr>
			<td>Bone morphogenetic protein 4 (BMP-4)</td>
			<td>PeproTech Inc.</td>
			<td>#120-05A</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine serum albumin (BSA)</td>
			<td>Sigma-Aldrich</td>
			<td>#A8022-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Cytokeratin 12 antibody</td>
			<td>Santa Cruz Biotechnology</td>
			<td>#SC-17099</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>Cytokeratin 14 antibody</td>
			<td>R&#38;D Systems</td>
			<td>#MAB3164</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>Cytokeratin 15 antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>#MS-1068-P</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>CnT-30</td>
			<td>CELLnTEC Advanced Cell Systems AG</td>
			<td>#Cnt-30</td>
			<td>Culture medium for adherent hPSC-LESC differentiation</td>
		</tr>
		<tr>
			<td>Collagen type IV (human)</td>
			<td>Sigma-Aldrich</td>
			<td>#C5533</td>
			<td>Human placental collagen type IV</td>
		</tr>
		<tr>
			<td>CoolCell LX Freezing Container</td>
			<td>Sigma-Aldrich</td>
			<td>#BCS-405</td>
			<td></td>
		</tr>
		<tr>
			<td>CryoPure tubes</td>
			<td>Sarsted</td>
			<td>#72.380</td>
			<td>1.6 ml cryotube for hPSC-LESC cryopreservation</td>
		</tr>
		<tr>
			<td>Defined Trypsin Inhibitor</td>
			<td>Gibco</td>
			<td>#R-007-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Essential 8 Flex Medium Kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>#A2858501</td>
			<td></td>
		</tr>
		<tr>
			<td>GlutaMAX</td>
			<td>Gibco</td>
			<td>#35050061</td>
			<td></td>
		</tr>
		<tr>
			<td>Laminin 521</td>
			<td>Biolamina</td>
			<td>#Ln521</td>
			<td>Human recombinant laminin 521</td>
		</tr>
		<tr>
			<td>&#916;Np63&#945; antibody</td>
			<td>BioCare Medical</td>
			<td>#4892</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>OCT3/4 antibody</td>
			<td>R&#38;D Systems</td>
			<td>#AF1759</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>p63&#945; antibody</td>
			<td>Cell Signaling Technology</td>
			<td>#ACI3066A</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>p63-&#945; (D2K8X) XP Rabbit mAb (PE Conjugate)</td>
			<td>Cell Signaling Technology</td>
			<td>#56687</td>
			<td>p63-&#945; PE-conjugated antibody for flow cytometry</td>
		</tr>
		<tr>
			<td>PAX6 antibody</td>
			<td>Sigma-Aldrich</td>
			<td>#HPA030775</td>
			<td>Primary antibody for IF</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Lonza</td>
			<td>#17-602E</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde (PFA)</td>
			<td>Sigma-Aldrich</td>
			<td>#158127</td>
			<td>Cell fixative for IF</td>
		</tr>
		<tr>
			<td>ProLong Gold Antifade Mountant with DAPI</td>
			<td>Thermo Fisher Scientific</td>
			<td>#P36931</td>
			<td>DAPI mountant for hard mounting for IF</td>
		</tr>
		<tr>
			<td>PSC Cryopreservation Kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>#A2644601</td>
			<td></td>
		</tr>
		<tr>
			<td>TrypLE Select Enzyme</td>
			<td>Gibco</td>
			<td>#12563-011</td>
			<td></td>
		</tr>
		<tr>
			<td>KnockOut Dulbecco&#8217;s modified Eagle&#8217;s medium</td>
			<td>Gibco</td>
			<td>#10829018</td>
			<td></td>
		</tr>
		<tr>
			<td>KnockOut SR XenoFree CTS</td>
			<td>Gibco</td>
			<td>#10828028</td>
			<td></td>
		</tr>
		<tr>
			<td>MEM non-essential amino acids</td>
			<td>Gibco</td>
			<td>#11140050</td>
			<td></td>
		</tr>
		<tr>
			<td>SB-505124</td>
			<td>Sigma-Aldrich</td>
			<td>#S4696</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma-Aldrich</td>
			<td>#T8787</td>
			<td>Permeabilization agent for IF</td>
		</tr>
		<tr>
			<td>VectaShield</td>
			<td>Vector Laboratories</td>
			<td>#H-1200</td>
			<td>DAPI mountant for liquid mounting for IF</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cytocentrifuge, e.g. CellSpin II</td>
			<td>Tharmac</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Flow cytometer, e.g. BD Accuri C6</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence microscope, e.g.Olympus IX 51</td>
			<td>Olympus</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

efficient and scalable directed differentiation of clinically compatible corneal limbal epithelial stem cells from human pluripotent stem cells

角膜缘上皮干细胞 (LESCs) 负责持续更新角膜上皮细胞, 从而保持角膜的稳态和视觉清晰度。人类多能干细胞 (hPSC) 衍生 LESCs 为角膜细胞置换治疗提供了有希望的细胞来源。未定义、异种的培养和分化条件导致了研究结果的变化, 阻碍了 hPSC 治疗的临床翻译。该协议为异种和馈线无细胞条件下的 hPSC LESC 分化提供了一种可重现和有效的方法。首先, 对重组 laminin-521 (LN-521) 和定义的 hPSC 培养基中未分化 hPSC 的单层培养是稳定生产高质量起始材料的基础。其次, 快速简单的 hPSC LESC 分化方法仅在24天内就会产生 LESC 种群。该方法包括四天表面外胚层诱导的小分子悬浮液, 其次是在 LN-521/collagen IV 组合基质上贴壁培养相, 确定角膜上皮分化培养基。Cryostoring 和扩展分化进一步净化细胞的种群, 并使细胞的银行在大量的细胞治疗产品。由此产生的高质量 hPSC LESCs 为角膜表面重建治疗角膜缘干细胞缺乏 (康乐文化署) 提供了一种潜在的新的处理策略。

从 hPSCs 到 hPSC-LESCs
从诱发 FF hPSCs 到 cryostoring hPSC LESCs 的整个过程大约需要3.5 周。图 1A给出了突出其关键步骤的微分方法的原理图概述。图 1B显示了协议不同阶段细胞种群的典型形态。所提供的数据采用 Regea08/017 人类胚胎干细胞线和 04607. WT hiPSC 线, 在芬兰坦佩雷大学获得和表征, 如?...

Watch this Scientific Journal Video about Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells at JoVE.com

Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells

高效可扩展的人多潜能干细胞临床相容角膜缘上皮干细胞的定向分化

本协议介绍了一种简单的两步法在异种和无细胞培养条件下, 从人多潜能干细胞中分化角膜缘上皮干细胞的方法。这里介绍的细胞培养方法可实现经济高效的大规模生产, 适用于角膜细胞治疗使用的临床质量细胞。

Corneal limbal epithelial stem cells (LESCs) are responsible for continuously renewing the corneal epithelium, and thus maintaining corneal homeostasis ...

该协议引入了一种定义的支线细胞自由方法，用于将角膜边缘上皮干细胞与人类多能干细胞进行区分。肢上皮干细胞负责在健康的眼睛中更新角膜上皮。这些细胞的损伤导致一种称为边缘干细胞缺乏症的情况，并导致角膜清晰度的丧失。此处所示的细胞培养方法能够有效生产肢体上皮干细胞，用于未来的角膜细胞替代疗法。首先，通过和维护文本协议中概述的免费人类多能干细胞培养。确保只使用高质量的人类多能干细胞作为分化的起始材料。当准备诱导胚胎体形成时，将所需的所有材料和试剂加热到层流罩中的室温。通过向每口井添加 500 微升的异种自由蛋白蛋白 EDTA，将喂食器自由人多能干细胞分离到悬浮液中。在37摄氏度下孵育，二氧化碳为5%。三分钟后，从培养箱中取出细胞，检查细胞形态，以确定去除三辛的最佳阶段。仔细观察细胞，以确定最佳时间去除异种自由尝试EDTA，当细胞已经四舍五入，但还没有完全分离。在最佳时间，从细胞中去除无异种的尝试素EDTA。添加定义的三辛抑制剂和轻轻移液器分离细胞。在50毫升离心管中收集单细胞悬浮液。在300g下离心5分钟。然后，去除上心，并在培养培养的一毫升中重新暂停细胞。数数细胞后，将两到三百万个细胞在三毫升基底诱导介质中分配，并辅以五微摩尔血红素，以低附六井板的每个井。在37摄氏度下孵育，二氧化碳为5%。第二天，在显微镜下检查胚胎体的质量。去除介质，代之以三毫升基底感应介质，辅以10微摩尔SB-505124和50毫微克每毫升人类基本玻璃纤维生长因子。继续在37摄氏度下孵育，二氧化碳为5%。之后两天，取出该介质，代之以三毫升的基础诱导介质，每毫升骨形态遗传蛋白四分之二五纳米。然后，使用以前的条件继续孵育。在第四天，准备每平方厘米的拉米宁-5210.5微克的混合物，以及每平方厘米5微克的人类胎盘胶原蛋白四型，在含有钙2和镁2离子的DPBS中稀释。使用这种混合物，涂覆100毫米组织培养皿，使粘附剂分化，总涂层体积为每盘5毫升。密封盘子，并储存在四摄氏度过夜。在第五天，将所有所需的材料和试剂加热到层流罩中的室温。在此之后，去除涂层溶液，并添加 10 毫升预热分化介质到每个菜。使用移液器，将胚胎体从一个盘子中移植到两到三个组织培养皿中。然后，轻轻摇动每个培养皿，均匀分布胚胎体。在未来两个半星期到三周内，在37摄氏度的粘附培养下保持细胞的二氧化碳，确保每周三次用10毫升新鲜分化介质代替该培养。使用相位对比显微镜定期检查细胞是否出现正确的上皮形态。首先，将所需的所有材料和试剂预热至层流罩中的室温，但应预冷冻的低温保存介质除外。接下来，通过加入无异种的三辛蛋白EDTA，在37摄氏度下以5%的二氧化碳孵育，将人类多能干细胞衍生的肢体上皮干细胞分离到单细胞悬浮液中。五分钟后，从培养箱中取出细胞并检查细胞形态，以确定去除三辛的最佳阶段。仔细观察细胞，以确定最佳时间，以删除异种自由尝试素EDTA，当细胞已经四舍五入，但尚未完全分离。在最佳时间，从细胞中去除异种自由尝试素 EDTA，并如前所述分离细胞。数数细胞后，以300g离心5分钟。吸气介质，并在预冷的异种自由冷冻保存介质中重新释放细胞。使用移液器，将单细胞悬浮液转移到低温管中，使每个冷冻管在一毫升低温保存介质中含有50万到100万个细胞。将管子放在冷冻容器中。在五分钟内，将它们转移到一个冷冻室在负80摄氏度过夜存储。第二天，将管子转移到液氮中长期储存。在解冻细胞之前，用每平方厘米五微克的人类胎盘胶原蛋白四型和每平方厘米 LM-521 的 0.5 微克的混合物，将所有所需的材料和试剂预热，使其达到层流罩中的室温。接下来，从盘子和板井中去除涂层溶液，并添加适当体积的预热分化介质。将预热分化介质添加到 15 毫升锥形管中。快速将细胞解冻至室温。解冻后，立即将电池悬浮液转移到锥形离心管。在300g下离心5分钟。吸气介质，并在分化介质中重新暂停细胞颗粒，以去除任何低温保存介质。以每平方厘米4万至5万个细胞的密度，将细胞镀在分型介质的预涂盘上。将细胞保持在37摄氏度的5%CO2下，每周更换三次分化介质。在Laminin-521上，未分化的高质量喂食器自由人类多能干细胞首先形成具有锋利的边缘的不同菌落，在汇合时合并成同质的单层。在基础诱导介质中，辅以5微摩尔血红蛋白24小时通常产生各种大小的紧密、常规胚胎体悬浮体，而胚胎体形态在悬浮液表面异体诱导期间不应发生显著变化，在胚胎体被镀到四型胶原蛋白和拉米宁-521组合基质中，发现殖民地外产会很快出现。在分化的21至25天内，细胞形成汇合均匀的均匀层，具有多边形形态，这是上皮细胞的典型特征。然后，这些细胞可以冷冻储存起来供以后使用，因为解冻后，生存能力和形态都保存完好。在分化的第24天，绝大多数细胞表示成对盒蛋白，PAX6，眼睛发育的关键调节器，以及p63 alpha，广泛认可的LESC标记。Delta Np63 在大多数 p63 alpha 阳性细胞中共同表达， 确认最角膜特异性增量 Np63 alpha 阳性细胞表型.其他标记部分表达，而其他标记在这一点上是检测不到的，表明分化已经进展到非全权边缘上皮祖细胞，但最终分化到成熟的角膜上皮细胞尚未发生。平均而言，每个未分化的喂食器自由人类多能干细胞在25日产生0.7个细胞。第24天，至少预计有65%的三角洲Np63α阳性细胞量。冷冻保存进一步净化细胞群体。总体而言，此处介绍的方法相对简单，但有一些成功的关键点。高品质的起始材料是必不可少的，以及温和，流利的细胞培养技术一般。该协议为临床应用和各种研究目的提供强大的方法，用于生产肢体上皮干细胞。此外，该方法可以很容易地修改，以分化视网膜色素上皮细胞。

Research

JoVE Journal

Developmental Biology

Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol

从人多能干细胞使用高度可重复的小分子为基础的分化协议心肌细胞规模化生产

Maximizing the benefit of human pluripotent stem cells (hPSCs) for research, disease modeling, pharmaceutical and clinical applications requires robust ...

科研

发育生物学

The Production of Pluripotent Stem Cells from Mouse Amniotic Fluid Cells Using a Transposon System

从小鼠羊水细胞多能干细胞使用转座子系统的生产

Induced pluripotent stem (iPS) cells are generated from mouse and human somatic cells by forced expression of defined transcription factors using ...

Directed Differentiation of Primitive and Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells

人多能干细胞对原始和最终造血祖细胞定向分化的研究

One of the major goals for regenerative medicine is the generation and maintenance of hematopoietic stem cells (HSCs) derived from human pluripotent stem ...

Efficient Differentiation of Pluripotent Stem Cells to NKX6-1+ Pancreatic Progenitors

Efficient Differentiation of Pluripotent Stem Cells to NKX6-1+ Pancreatic Progenitors

多能高效分化干细胞NKX6-1<sup&gt; +</sup&gt;胰腺祖

Pluripotent stem cells have the ability to self renew and differentiate to multiple lineages, making them an attractive source for the generation of ...

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

在人多能的体外干细胞向滋养层细胞

The placenta is the first organ to develop during embryogenesis and is required for the survival of the developing embryo. The placenta is comprised of ...

Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells

界定和由人多能干细胞的肝细胞样细胞的可扩展代

Human pluripotent stem cells (hPSCs) possess great value for biomedical research. hPSCs can be scaled and differentiated to all cell types found in the ...

Rapid, Directed Differentiation of Retinal Pigment Epithelial Cells from Human Embryonic or Induced Pluripotent Stem Cells

人胚胎或诱导多能干细胞对视网膜色素上皮细胞的快速定向分化

We describe a robust method to direct the differentiation of pluripotent stem cells into retinal pigment epithelial cells (RPE). The purpose of providing ...

Efficient Differentiation of Human Pluripotent Stem Cells into Liver Cells

人类多能干细胞有效分化为肝细胞

The liver detoxifies harmful substances, secretes vital proteins, and executes key metabolic activities, thus sustaining life. Consequently, liver ...

In Vitro Generation of Somite Derivatives from Human Induced Pluripotent Stem Cells

人诱导多能干细胞体外生成体细胞衍生物的研究

In response to signals such as WNTs, bone morphogenetic proteins (BMPs), and sonic hedgehog (SHH) secreted from surrounding tissues, somites (SMs) give ...

Transient Treatment of Human Pluripotent Stem Cells with DMSO to Promote Differentiation

用DMSO对人多能干细胞的暂时性治疗促进分化

Despite the growing use of pluripotent stem cells (PSCs), challenges in efficiently differentiating embryonic and induced pluripotent stem cells (ESCs and ...