Brain pericytes regulate diverse aspects of neurovascular development and function, including blood-brain barrier (BBB) induction and maintenance. Primary brain pericytes have been widely employed in coculture-based in vitro models of the BBB, and a method to generate brain pericytes from human pluripotent stem cells (hPSCs) could provide a renewable, genetically tractable source of cells for BBB modeling and studying pericyte roles in development and disease. Here, we describe a protocol to differentiate hPSCs to NG2+ PDGFRβ+ αSMAlow brain pericyte-like cells in 22-25 days through a p75-NGFR+ HNK-1+ neural crest intermediate, which mimics the developmental origin of forebrain pericytes. The resulting brain pericyte-like cells have molecular and functional attributes of brain pericytes. We also provide protocols for maintenance, cryopreservation, and recovery of the neural crest intermediate, and for molecular and functional characterization of the resulting cells.
Brain pericytes regulate diverse aspects of neurovascular development and function, including blood-brain barrier (BBB) induction and maintenance. Primary brain pericytes have been widely employed in coculture-based in vitro models of the BBB, and a method to generate brain pericytes from human pluripotent stem cells (hPSCs) could provide a renewable, genetically tractable source of cells for BBB modeling and studying pericyte roles in development and disease. Here, we describe a protocol to differentiate hPSCs to NG2+ PDGFRβ+ αSMAlow brain pericyte-like cells in 22-25 days through a p75-NGFR+ HNK-1+ neural crest intermediate, which mimics the developmental origin of forebrain pericytes. The resulting brain pericyte-like cells have molecular and functional attributes of brain pericytes. We also provide protocols for maintenance, cryopreservation, and recovery of the neural crest intermediate, and for molecular and functional characterization of the resulting cells.
Authors: Axel Montagne; Angeliki M Nikolakopoulou; Zhen Zhao; Abhay P Sagare; Gabriel Si; Divna Lazic; Samuel R Barnes; Madelaine Daianu; Anita Ramanathan; Ariel Go; Erica J Lawson; Yaoming Wang; William J Mack; Paul M Thompson; Julie A Schneider; Jobin Varkey; Ralf Langen; Eric Mullins; Russell E Jacobs; Berislav V Zlokovic Journal: Nat Med Date: 2018-02-05 Impact factor: 53.440
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Authors: Ross Nortley; Nils Korte; Pablo Izquierdo; Chanawee Hirunpattarasilp; Anusha Mishra; Zane Jaunmuktane; Vasiliki Kyrargyri; Thomas Pfeiffer; Lila Khennouf; Christian Madry; Hui Gong; Angela Richard-Loendt; Wenhui Huang; Takashi Saito; Takaomi C Saido; Sebastian Brandner; Huma Sethi; David Attwell Journal: Science Date: 2019-06-20 Impact factor: 47.728