Name: efficient differentiation of human pluripotent stem cells into liver cells
Uploaded: 2019-06-11T13:30:00
Description: Watch this Scientific Journal Video about Efficient Differentiation of Human Pluripotent Stem Cells into Liver Cells at JoVE.com

我们感谢 Bing Lim 的讨论,以及斯坦福干细胞生物学和再生医学研究所的基础设施支持。这项工作得到了加州再生医学研究所(DISC2-10679)和斯坦福大学-伯克利西贝尔干细胞研究所(L.T.A.和K.M.L.)和斯坦福贝克曼分子和基因医学中心以及匿名组织的支持。巴克斯特和迪杰诺娃家族(到K.M.L.)。

1. 差异化介质的准备
注:有关所用材料和试剂的制造商信息,请参阅材料表。
<ol><li>
制备基础化学定义介质 注:CDM2、CDM3、CDM4和CDM5是化学定义的介质,用作不同阶段将hPSC与肝细胞区分为基介质的基础介质。这些介质的组成可在表1中找到。<ol>
<li>要制作 CDM2 或 CDM3,请准备含有聚乙烯醇 (PVA) 的库存溶液。将 0.5 g 的 PVA 粉末溶解...

<ol>
	<li>Bernal, W., Wendon, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+Liver+Failure.">Acute Liver Failure.</a> New England Journal of Medicine. 369, 2525-2534 (2013).</li><li>Ang, L. T., 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=A+Roadmap+for+Human+Liver+Differentiation+from+Pluripotent+Stem+Cells.">A Roadmap for Human Liver Differentiation from Pluripotent Stem Cells.</a> Cell Reports. 22, 2190-2205 (2018).</li><li>Fisher, R. A., Strom, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+Hepatocyte+Transplantation:+Worldwide+Results.">Human Hepatocyte Transplantation: Worldwide Results.</a> Transplantation. 82, 441-449 (2006).</li><li>Dhawan, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+human+hepatocyte+transplantation:+Current+status+and+challenges.">Clinical human hepatocyte transplantation: Current status and challenges.</a> Liver transplantation. 21, S39-S44 (2015).</li><li>Salama, H., 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=Autologous+Hematopoietic+Stem+Cell+Transplantation+in+48+Patients+with+End-Stage+Chronic+Liver+Diseases.">Autologous Hematopoietic Stem Cell Transplantation in 48 Patients with End-Stage Chronic Liver Diseases.</a> Cell Transplantation. 19, 1475-1486 (2017).</li><li>Tan, A. K. Y., Loh, K. M., Ang, L. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+the+regenerative+potential+and+functionality+of+human+liver+cells+in+mice.">Evaluating the regenerative potential and functionality of human liver cells in mice.</a> Differentiation. 98, 25-34 (2017).</li><li>Loh, K. M., 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+Endoderm+Induction+from+Human+Pluripotent+Stem+Cells+by+Logically+Directing+Signals+Controlling+Lineage+Bifurcations.">Efficient Endoderm Induction from Human Pluripotent Stem Cells by Logically Directing Signals Controlling Lineage Bifurcations.</a> Cell Stem Cell. 14, 237-252 (2014).</li><li>Loh, K. M., 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=Mapping+the+Pairwise+Choices+Leading+from+Pluripotency+to+Human+Bone,+Heart,+and+Other+Mesoderm+Cell+Types.">Mapping the Pairwise Choices Leading from Pluripotency to Human Bone, Heart, and Other Mesoderm Cell Types.</a> Cell. , 451-467 (2016).</li><li>Zhao, D., 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=Promotion+of+the+efficient+metabolic+maturation+of+human+pluripotent+stem+cell-derived+hepatocytes+by+correcting+specification+defects.">Promotion of the efficient metabolic maturation of human pluripotent stem cell-derived hepatocytes by correcting specification defects.</a> Cell Research. 23, 157-161 (2012).</li><li>Si Tayeb, K., 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=Highly+efficient+generation+of+human+hepatocyte-like+cells+from+induced+pluripotent+stem+cells.">Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells.</a> Hepatology. 51, 297-305 (2010).</li><li>Carpentier, A., 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=Hepatic+differentiation+of+human+pluripotent+stem+cells+in+miniaturized+format+suitable+for+high-throughput+screen.">Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen.</a> Stem Cell Research. 16, 640-650 (2016).</li><li>Avior, Y., 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=Microbial-derived+lithocholic+acid+and+vitamin+K2+drive+the+metabolic+maturation+of+pluripotent+stem+cells-derived+and+fetal+hepatocytes.">Microbial-derived lithocholic acid and vitamin K2 drive the metabolic maturation of pluripotent stem cells-derived and fetal hepatocytes.</a> Hepatology. 62, 265-278 (2015).</li><li>Ogawa, S., 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=Three-dimensional+culture+and+cAMP+signaling+promote+the+maturation+of+human+pluripotent+stem+cell-derived+hepatocytes.">Three-dimensional culture and cAMP signaling promote the maturation of human pluripotent stem cell-derived hepatocytes.</a> Development. 140, 3285-3296 (2013).</li><li>Rostovskaya, M., Bredenkamp, N., Smith, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+consistent+generation+of+pancreatic+lineage+progenitors+from+human+pluripotent+stem+cells.">Towards consistent generation of pancreatic lineage progenitors from human pluripotent stem cells.</a> Philosophical Transactions of the Royal Society B: Biological Sciences. 370, 20140365 (2015).</li><li>Haridass, D., 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=Repopulation+Efficiencies+of+Adult+Hepatocytes,+Fetal+Liver+Progenitor+Cells,+and+Embryonic+Stem+Cell-Derived+Hepatic+Cells+in+Albumin-Promoter-Enhancer+Urokinase-Type+Plasminogen+Activator+Mice.">Repopulation Efficiencies of Adult Hepatocytes, Fetal Liver Progenitor Cells, and Embryonic Stem Cell-Derived Hepatic Cells in Albumin-Promoter-Enhancer Urokinase-Type Plasminogen Activator Mice.</a> The American Journal of Pathology. 175, 1483-1492 (2009).</li></ol>

这种方法使丰富的肝芽祖细胞,以及随后的肝细胞样细胞,从hPSC生成。产生丰富人类肝细胞种群的能力对于实际利用这种细胞非常重要。以前从hPSCs生成肝细胞的方法产生了含有肝脏和非肝细胞的不纯细胞群,在移植到啮齿动物体内后,除了肝组织15外,还产生了骨骼和软骨。因此,明确抑制非肝分化对于产生可能适合各种应用的丰富肝脏种群至关重要。
值得注意的...

该协议的目的是有效地区分人类多能干细胞(hPSCs)到丰富的肝脏芽祖细胞和肝细胞样细胞2。获得现成的人类肝脏祖和肝细胞样细胞将加速研究肝功能和疾病的努力,并可能支持新的细胞移植疗法肝衰竭3,4 5.这在过去被证明是具有挑战性的,因为hPSCs(包括胚胎和诱导多能干细胞)可以分化成人体的所有细胞类型;因此,很难将它们完全区分为单细胞类型的纯种群,如肝细胞6。
要将hPSC精确区分为肝细胞,首先不仅要了解肝细胞是如何指定的,还要了解非肝细胞类型是如何发育的。了解非肝细胞如何发育对于在分化期间从逻辑上抑制非肝系的形成非常重要,从而专门指导hPSC走向肝脏命运2。其次,必须划定hPSC区分肝脏命运的多重发育步骤。众所周知,hPSCs在形成肝细胞样细胞(HEP)之前,会按顺序分化成多种细胞类型,称为原始条纹(APS)、最终内皮(DE)、后前体(PFG)和肝芽祖细胞(LB)。早期的工作揭示了指示肝脏命运的信号和抑制交替非肝细胞类型(包括胃、胰腺和肠道祖细胞)形成的信号,每个发育系选择2,7,8.
总体而言,这些见解已经产生了一种无血清,单层方法,以区分hPSCs对原始条纹,明确的内皮,后前体,肝芽祖,最后,肝细胞样细胞2。总体而言,该方法涉及以适当的密度在单层中播种 hPSC,制备六种分化介质的鸡尾酒(包含调节各种发育信号通路的生长因子和小分子),以及依次添加这些介质,在18天内诱导分化。在此过程中,不需要细胞的传递。值得注意的是,由于这种方法明确包括抑制非肝细胞类型形成的信号,这种分化方法1通过与现存的肝细胞和肝细胞样细胞的比较,更有效地生成肝祖和肝细胞样细胞区分方法2,9,10,11,12 。此外,本文中描述的协议使肝细胞的生成更快,最终表达出比其他协议9、10产生的肝转录因子和酶的更高水平,11,12.
与当前差别化协议不同,此处描述的协议具有某些优势。首先,它需要hPSC的单层分化,这在技术上比三维分化方法要简单,例如那些依赖于胚胎体的方法13。其次,这种方法利用了最近的进步,即最终内皮细胞(肝细胞的早期前体)可以在hPSC分化2,7的2天内有效和迅速生成,从而使后续生产纯度提高的肝细胞。第三,在并排比较中,该方法2产生的肝细胞样细胞产生更多的ALBUMIN,表达出比其它方法产生的肝细胞更高水平的肝转录因子和酶。 11,12.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr >
			<td >&#160;Geltrex</td>
			<td >Thermofisher Scientific</td>
			<td >A1569601</td>
			<td ></td>
		</tr>
		<tr >
			<td >1:1 DMEM/F12</td>
			<td>Gibco</td>
			<td>11320033</td>
			<td></td>
		</tr>
		<tr >
			<td >0.2&#160;&#956;m pore membrane filter</td>
			<td>Millipore</td>
			<td>GTTP02500</td>
			<td ></td>
		</tr>
		<tr >
			<td >mTeSR1</td>
			<td>Stem Cell Technologies</td>
			<td>5850</td>
			<td></td>
		</tr>
		<tr >
			<td >Thiazovivin</td>
			<td>Tocris Bioscience</td>
			<td>3845</td>
			<td></td>
		</tr>
		<tr >
			<td >&#160;Accutase</td>
			<td>Gibco or Millipore&#160;</td>
			<td>Gibco A11105, Millipore&#160; SCR005</td>
			<td></td>
		</tr>
		<tr >
			<td >IMDM, GlutaMAX&#8482; Supplement</td>
			<td>Thermofisher Scientific</td>
			<td>31980030</td>
			<td></td>
		</tr>
		<tr >
			<td >Ham&#39;s F-12 Nutrient Mix, GlutaMAX&#8482; Supplement</td>
			<td>Thermofisher Scientific</td>
			<td>31765035</td>
			<td></td>
		</tr>
		<tr >
			<td >KOSR, Knockout serum replacement</td>
			<td>Thermofisher Scientific</td>
			<td>10828028</td>
			<td></td>
		</tr>
		<tr >
			<td >Poly(vinyl alcohol)</td>
			<td>Sigma-Aldrich&#160;&#160;</td>
			<td>P8136</td>
			<td></td>
		</tr>
		<tr >
			<td >Transferrin&#160;</td>
			<td >Sigma-Aldrich&#160;&#160;</td>
			<td >10652202001</td>
			<td></td>
		</tr>
		<tr >
			<td >Chemically Defined Lipid Concentrate</td>
			<td>Thermofisher Scientific</td>
			<td >11905031</td>
			<td></td>
		</tr>
		<tr >
			<td >human Activin</td>
			<td >R&#38;D</td>
			<td>338-AC</td>
			<td></td>
		</tr>
		<tr >
			<td >CHIR99201</td>
			<td >Tocris</td>
			<td>4423</td>
			<td></td>
		</tr>
		<tr >
			<td >PI103</td>
			<td >Tocris</td>
			<td >2930/1</td>
			<td></td>
		</tr>
		<tr >
			<td >human FGF2</td>
			<td >R&#38;D</td>
			<td>233-FB</td>
			<td></td>
		</tr>
		<tr >
			<td >DM3189</td>
			<td >Tocris</td>
			<td>6053/10</td>
			<td></td>
		</tr>
		<tr >
			<td >A83-01</td>
			<td >Tocris</td>
			<td>2939/10</td>
			<td></td>
		</tr>
		<tr >
			<td >Human BMP4</td>
			<td >R&#38;D</td>
			<td>314-BP</td>
			<td></td>
		</tr>
		<tr >
			<td >C59</td>
			<td >Tocris</td>
			<td>5148</td>
			<td></td>
		</tr>
		<tr >
			<td >TTNPB</td>
			<td >Tocris</td>
			<td>0761/10</td>
			<td></td>
		</tr>
		<tr >
			<td >Forskolin</td>
			<td >Tocris</td>
			<td>1099/10</td>
			<td></td>
		</tr>
		<tr >
			<td >Oncostatin M</td>
			<td >R&#38;D</td>
			<td>295-OM</td>
			<td></td>
		</tr>
		<tr >
			<td >Dexamethasone</td>
			<td >Tocris</td>
			<td>1126</td>
			<td></td>
		</tr>
		<tr >
			<td >Ro4929097</td>
			<td >Selleck Chem</td>
			<td>S1575</td>
			<td></td>
		</tr>
		<tr >
			<td >AA2P</td>
			<td>Cayman chemicals</td>
			<td>16457</td>
			<td></td>
		</tr>
		<tr >
			<td >human recombinant Insulin</td>
			<td>Sigma-Aldrich&#160;&#160;</td>
			<td>11061-68-0</td>
			<td></td>
		</tr>
	</tbody>
</table>

efficient differentiation of human pluripotent stem cells into liver cells

肝脏排毒有害物质,分泌重要蛋白质,并执行关键的代谢活动,从而维持生命。因此,肝衰竭(可能由慢性酒精摄入、肝炎、急性中毒或其他侮辱引起)是一种严重疾病,可最终导致出血、黄热病、昏迷,并最终死亡。然而,治疗肝功能的方法,以及肝功能和疾病的研究,由于人类肝细胞的缺乏,部分受阻。为此,该协议详细介绍了人类多能干细胞(hPSCs)与肝细胞样细胞的有效分化,其指导是一个发展路线图,描述了肝脏命运是如何在六个连续分化步骤中指定的。通过操纵发育信号通路,促进肝脏分化,并明确抑制不需要的细胞命运的形成,这种方法在第6天有效地生成人类肝芽祖细胞和肝细胞样细胞的种群和 PSC 差异的 18 个。这是通过时间精确控制发展信号通路实现的,由无血清培养基中的小分子和生长因子施加。该系统的分化发生在单层,并产生肝细胞样细胞,表达特征肝细胞酶,并有能力移植慢性肝衰竭的小鼠模型。在体外有效生成大量人类肝细胞的能力对治疗肝衰竭、药物筛选和肝病的机械研究都有影响。

在 APS 分化 24 小时后,菌落通常采用不同于未分化的菌落的形态,同时失去通常限制 hPSC 菌落的明亮边界。形态上,原始条纹细胞通常具有不规则的边界,比hPSCs传播更紧密,更紧凑-这是令人回味的上皮到中生体过渡,因为多能表细胞分化和进入原始条纹在体内。如果分化前的hPSC的菌落大小过大,将有一个明显升高的中心,由未分化的细胞组成。含有如此大团状的培养物应该被丢弃,...

Watch this Scientific Journal Video about Efficient Differentiation of Human Pluripotent Stem Cells into Liver Cells at JoVE.com

Efficient Differentiation of Human Pluripotent Stem Cells into Liver Cells

该协议详细介绍了单层无血清方法,在18天内从人类多能干细胞(hPSCs)中高效生成肝细胞样细胞。这需要六个步骤,因为hPSCs在形成肝细胞样细胞之前,按顺序分化成中间细胞类型,如原始条纹、最终内皮、后前体和肝芽祖细胞。

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 ...

这种方法能够产生大量的人类肝芽祖细胞和高纯度的肝细胞样细胞。产生这些肝细胞的能力对于再生医学和其他领域可能很重要。通过控制发育信号通路，促进肝脏分化，抑制不需要的细胞类型的形成，这种方法使人类肝芽祖细胞和肝细胞样细胞分别有效生成6天和18天的分化。这项技术的主要优点是产生的人类肝细胞纯度高。要执行此过程，请将 50 毫升 IMDM 添加到包含搅拌棒的圆锥烧瓶中。加热到50摄氏度的介质，加入0.5克PVA，同时连续搅拌，以每毫升10毫克的浓度准备PVA库存。在此之后，从加热垫上取出 PVA 溶液，让其冷却至室温。冷却溶液后，通过无菌的 0.2 微米过滤器进行过滤。通过将过滤的PVA与其他试剂结合，准备CDM2和CDM3，如文本协议表之一所述。使用无菌 0.2 微米过滤器过滤所有介质。然后，编写其余基本介质，如案文议定书表一所述，清洁发展机制4和清洁发展机制5。要准备分化介质，首先在室温下解冻冷冻小分子和/或生长因子。将所需量的基介质分出，并在适当的浓度下将指定的小分子和生长因子添加到基介质中。使用前一天，在摄氏四度时解冻矩阵。第二天，将500微升基质加入50毫升冷DMEM，以1比100的比例稀释基质。使用足够体积的溶液将稀释的基体移入所需数量的油井中，以覆盖油井表面。将基质涂层板转移到 37 摄氏度的培养箱中至少 60 分钟。在播种之前，立即从涂层孔中吸出剩余的基质溶液。然后，从大部分汇合的 hPSC 培养板中吸出介质，并添加商业购买的分离剂，确保仅使用足够的介质来覆盖细胞生长的表面。在37摄氏度下孵育分离剂中的hPSCs5分钟，或直到一些菌落开始分离。接下来，添加DMEM和F-12以稀释分离剂。使用五毫升血清移液器多次轻轻上下移液器，以洗掉井表面的所有细胞。在50毫升锥形管中收集重新暂停的单细胞，并根据需要用DMEM和F-12稀释。将收集的hPSCs管在350倍g和4摄氏度下离心3分钟，以对细胞进行分粒。在等待离心时，从将播种 hPSC 的板中吸出矩阵。向接收井添加足够数量的 mTeSR 介质以覆盖它们。离心完成后，小心吸进上清液，将颗粒的 hPSC 留在锥形管的底部。在 mTeSR 中重新补充细胞颗粒，并辅以商业获得的蒂亚佐维文的微摩尔。使用P1000移液器，轻轻三重搅拌两到三次，均匀地将细胞颗粒重新悬浮到单个细胞悬浮液中。立即移液10微升细胞悬浮成血细胞计并计算细胞数量。使用硫酮补充的 mTeSR 调节重新消耗的 hPSCs 的体积，以实现电镀所需的细胞浓度。然后，以交叉图案摇动板几次，以确保细胞均匀分布。在hPSC镀上至少24小时后，使用相位对比显微镜检查细胞的形态，并特别强调镀hPSC菌落的直径。接下来，准备第一天 APS 差异化介质，如文本协议表二所示。播种后一天，从镀制的 hPSC 中吸出蒂亚佐维文补充的 mTeSR，然后用 IMDM 介质短暂清洗。在此之后，向 hPSC 添加第一天介质。记录时间，将细胞放回37摄氏度的培养箱。继续后续的分化步骤，准备所需的分化介质，并在一天中同一时间将该介质添加到各自的分化日。在这项研究中，丰富的肝芽祖细胞和随后的肝细胞样细胞由hPSC产生。到分化的第二天，原始条纹细胞已经分化成第二天明确的内皮细胞。这些细胞中绝大多数表示SOX17和FOXA2。到分化的第三天，内端细胞已经分化成前体祖细胞，在形状上出现多边形。后来，到了第六天，前祖人分化成肝芽祖体。这些肝芽祖体表达 AFP、TPX3 和 HNF4alpha。在三条hPSC生产线上，这种方法产生第六天 AFP 阳性肝祖细胞，效率约为 89% 最后，在与 hPSC 的 18 天肝脏分化后，会出现白蛋白阳性的肝细胞样细胞。从形态学上来说，这些细胞出现上皮，形成明亮的边界，让人联想到胆汁。在这个阶段，18 hPSC衍生的肝细胞的细胞质看起来比核暗。当播种人类多能干细胞进行分化时，必须以正确的密度播种它们，并且通过摇动板将它们分布在井中。在体外产生人类肝细胞的能力是一种使能技术，使科学家能够研究肝脏发育的基础机制。这种方法在体外产生大量的人类肝细胞，可用于研究肝功能和疾病，并最终为肝衰竭提供新的疗法。

Research

JoVE Journal

Developmental Biology

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 ...

科研

发育生物学

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 ...

Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing

将人诱导的多能干细胞分化为神经元和神经胶质的混合培养物进行神经毒性测试的方案

Human pluripotent stem cells can differentiate into various cell types that can be applied to human-based in vitro toxicity assays. One major advantage is ...

In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells

In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells

在体外人骨髓间充质干细胞分化为心肌样细胞功能

Myocardial infarction and the subsequent ischemic cascade result in the extensive loss of cardiomyocytes, leading to congestive heart failure, the leading ...

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 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

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

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 ...

Efficient Neural Differentiation using Single-Cell Culture of Human Embryonic Stem Cells

利用人类胚胎干细胞的单细胞培养进行高效神经分化

In vitro differentiation of human embryonic stem cells (hESCs) has transformed the ability to study human development on both biological and molecular ...