Generation of Human Pluripotent Stem Cell-Derived Polarized Hepatocytes.
Generation of Human Pluripotent Stem Cell-Derived Polarized Hepatocytes.
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DOI：
10.1002/cpz1.345
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发表时间：
2022-01
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影响因子：
--
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中科院分区：
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人多能干细胞（hPSC）来源的肝细胞样细胞（HLC）是研究肝脏生物学的有价值的工具。然而，HLC缺乏与其生理功能相关的某些关键体内特征。其中一个特征是细胞极性，这对于涉及小管胆汁分泌和大量血清蛋白质窦状分泌到血液中的肝细胞逆流流动系统至关重要。因此，使用非极化肝细胞的模型系统不能概括肝细胞的这种生理功能。在这里，我们描述了一个逐步的协议，以产生hPSC衍生的极化的HLC（pol-HLC），其功能明确定义的基底外侧和顶端膜分离的紧密连接。Pol-HLC不仅显示许多肝脏功能，而且还能够定向分泌货物，模仿逆流流动系统。我们描述了用于干细胞培养维持和用于使hPSC分化成pol-HLC的方案。此外，我们还描述了检测pol-HLC的基本肝功能和极化肝脏特征的方案。一旦成功分化，这些pol-HLC可以用作体外模型系统来研究肝细胞生物学、疾病机制、遗传变异和药物代谢。从非极化的hPSC建立肝极性也提供了研究肝极性的发展和维持的有用工具。
Human pluripotent stem cells (hPSCs)-derived hepatocyte-like cells (HLCs) are valuable tools to study liver biology. HLCs, however, lack certain key in vivo characteristics relevant to their physiological function. One such characteristic is cellular polarity, which is critical to hepatocyte counter-current flow systems involving canalicular bile secretion and sinusoidal secretion of large quantities of serum proteins into blood. Model systems using non-polarized hepatocytes, therefore, cannot recapitulate this physiological function of hepatocytes. Here, we describe a stepwise protocol to generate hPSCs-derived polarized HLCs (pol-HLCs), which feature clearly defined basolateral and apical membranes separated by tight junctions. Pol-HLCs not only display many hepatic functions but are also capable of directional cargo secretion, mimicking the counter-current flow systems. We describe protocols for stem cell culture maintenance and for differentiating hPSCs into pol-HLCs. In addition, we describe protocols to assay the pol-HLCs for basic hepatic functions and polarized hepatic characteristics. Once successfully differentiated, these pol-HLCs can be used as an in vitro model system to study hepatocyte biology, disease mechanisms, genetic variation, and drug metabolism. The establishment of hepatic polarity from non-polarized hPSCs also provides a useful tool to study the development and maintenance of hepatic polarity.
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影响因子：
6.1
作者：
Godoy, Patricio;Hewitt, Nicola J.;Albrecht, Ute;Andersen, Melvin E.;Ansari, Nariman;Bhattacharya, Sudin;Bode, Johannes Georg;Bolleyn, Jennifer;Borner, Christoph;Boettger, Jan;Braeuning, Albert;Budinsky, Robert A.;Burkhardt, Britta;Cameron, Neil R.;Camussi, Giovanni;Cho, Chong-Su;Choi, Yun-Jaie;Rowlands, J. Craig;Dahmen, Uta;Damm, Georg;Dirsch, Olaf;Teresa Donato, Maria;Dong, Jian;Dooley, Steven;Drasdo, Dirk;Eakins, Rowena;Ferreira, Karine Sa;Fonsato, Valentina;Fraczek, Joanna;Gebhardt, Rolf;Gibson, Andrew;Glanemann, Matthias;Goldring, Chris E. P.;Jose Gomez-Lechon, Maria;Groothuis, Geny M. M.;Gustavsson, Lena;Guyot, Christelle;Hallifax, David;Hammad, Seddik;Hayward, Adam;Haeussinger, Dieter;Hellerbrand, Claus;Hewitt, Philip;Hoehme, Stefan;Holzhuetter, Hermann-Georg;Houston, J. Brian;Hrach, Jens;Ito, Kiyomi;Jaeschke, Hartmut;Keitel, Verena;Kelm, Jens M.;Park, B. Kevin;Kordes, Claus;Kullak-Ublick, Gerd A.;LeCluyse, Edward L.;Lu, Peng;Luebke-Wheeler, Jennifer;Lutz, Anna;Maltman, Daniel J.;Matz-Soja, Madlen;McMullen, Patrick;Merfort, Irmgard;Messner, Simon;Meyer, Christoph;Mwinyi, Jessica;Naisbitt, Dean J.;Nussler, Andreas K.;Olinga, Peter;Pampaloni, Francesco;Pi, Jingbo;Pluta, Linda;Przyborski, Stefan A.;Ramachandran, Anup;Rogiers, Vera;Rowe, Cliff;Schelcher, Celine;Schmich, Kathrin;Schwarz, Michael;Singh, Bijay;Stelzer, Ernst H. K.;Stieger, Bruno;Stoeber, Regina;Sugiyama, Yuichi;Tetta, Ciro;Thasler, Wolfgang E.;Vanhaecke, Tamara;Vinken, Mathieu;Weiss, Thomas S.;Widera, Agata;Woods, Courtney G.;Xu, Jinghai James;Yarborough, Kathy M.;Hengstler, Jan G.
通讯作者：
Hengstler, Jan G.
影响因子：
17.1
作者：
Liu H;Kim Y;Sharkis S;Marchionni L;Jang YY
通讯作者：
Jang YY
影响因子：
5.2
作者：
Hay, David C.;Zhao, Debiao;Cui, Wei
通讯作者：
Cui, Wei
影响因子：
--
作者：
WOOD, RL
通讯作者：
WOOD, RL
DOI：
10.1097/qai.0b013e3181eb376b
发表时间：
2010-11-01
影响因子：
3.6
作者：
German, Polina;Warren, David;Kearney, Brian P.
通讯作者：
Kearney, Brian P.