Generation of Brain Microvascular Endothelial-Like Cells from Human Induced Pluripotent Stem Cells by Co-Culture with C6 Glioma Cells

• Published in: PloS one 2015-06, Vol.10 (6), p.e0128890
• Main Authors: Minami, Haruka, Tashiro, Katsuhisa, Okada, Atsumasa, Hirata, Nobue, Yamaguchi, Tomoko, Takayama, Kazuo, Mizuguchi, Hiroyuki, Kawabata, Kenji
• Format: Article
• Language: English
• Subjects: Analysis; Animal models; Biochemistry; Blood; Blood-brain barrier; Brain; Brain - blood supply; Brain Neoplasms - pathology; Brain research; Cell culture; Cell Differentiation; Cell Line, Tumor; Coculture Techniques; Conditioning; Culture Media, Conditioned; Culture Media, Serum-Free; Dextran; Dextrans; Differentiation; Drug screening; Endothelial cells; Endothelium; Gene expression; Glioma; Glioma - pathology; Glioma cells; Gliomas; Humans; Induced Pluripotent Stem Cells - cytology; Inhibitory postsynaptic potentials; Laboratories; Medical research; Metabolism; Microvasculature; Molecular biology; Molecular chains; Neurological diseases; Neurophysiology; Permeability; Pharmaceutical sciences; Pluripotency; Stem cell research; Stem cell transplantation; Stem cells; Transport; Wnt protein; Wnt Signaling Pathway
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0128890

The blood brain barrier (BBB) is formed by brain microvascular endothelial cells (BMECs) and tightly regulates the transport of molecules from blood to neural tissues. In vitro BBB models from human pluripotent stem cell (PSCs)-derived BMECs would be useful not only for the research on the BBB development and function but also for drug-screening for neurological diseases. However, little is known about the differentiation of human PSCs to BMECs. In the present study, human induced PSCs (iPSCs) were differentiated into endothelial cells (ECs), and further maturated to BMECs. Interestingly, C6 rat glioma cell-conditioned medium (C6CM), in addition to C6 co-culture, induced the differentiation of human iPSC-derived ECs (iPS-ECs) to BMEC-like cells, increase in the trans-endothelial electrical resistance, decreased in the dextran transport and up-regulation of gene expression of tight junction molecules in human iPS-ECs. Moreover, Wnt inhibitors attenuated the effects of C6CM. In summary, we have established a simple protocol of the generation of BMEC-like cells from human iPSCs, and have demonstrated that differentiation of iPS-ECs to BMEC-like cells is induced by C6CM-derived signals, including canonical Wnt signals.