Human pluripotent stem cells (hPSCs) have emerged as an important source for cell therapy. However, to date, no studies demonstrated generation of purified hPSC-derived lymphatic endothelial cells (LECs) and tested their therapeutic potential in disease models. Here we sought to differentiate hPSCs into the LEC lineage, purify them with LEC markers, and evaluate their therapeutic effects. We found that an OP9-assisted culture system reinforced by addition of VEGF-A, VEGF-C, and EGF most efficiently generated LECs, which were then isolated via FACS-sorting with LYVE-1 and PODOPLANIN. These hPSC-derived LYVE-1 + PODOPLANIN + cells showed a pure committed LEC phenotype, formed new lymphatic vessels, and expressed lymphangiogenic factors at high levels. These hPSC-derived LECs enhanced wound healing through lymphangiogenesis and lymphvasculogenesis. Here we report, for the first time, that LECs can be selectively isolated from differentiating hPSCs, and that these cells are potent for lymphatic vessel formation in vivo and wound healing. This system and the purified hPSC-derived LECs can serve as a new platform for studying LEC development as well as for cell therapy.
Bibliographical noteFunding Information:
We thank Drs. George Daley and In-Hyun Park for providing the human iPSC line BJ1 and technical advice. This work was supported in part by the NIH grant DP3DK094346, HL119291, by the NHLBI of NIH as a Program of Excellence in Nanotechnology (HHSN268201000043C), the National Science Foundation STC grant CBET-0939511 and ACTSI pilot grant (PHS grant UL1 RR025008 from the CTSA program, NIH, and National Center for Research Resources).
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