Journal
STEM CELL RESEARCH & THERAPY
Volume 8, Issue -, Pages -Publisher
BIOMED CENTRAL LTD
DOI: 10.1186/s13287-016-0455-4
Keywords
Human pluripotent stem cells; Endothelial cells; MESP1; 3D culture; Vascularization
Funding
- National Basic Research Program of China [2012CB966701]
- National Natural Science Foundation of China (NSFC) [31171381, 51273106]
- Tsinghua University Initiative Scientific Research Program [20141081264]
- Tsinghua-Peking Center for Life Sciences
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Background: Generation of large quantities of endothelial cells is highly desirable for vascular research, for the treatment of ischemia diseases, and for tissue regeneration. To achieve this goal, we developed a simple, chemically defined culture system to efficiently and rapidly differentiate endothelial cells from human pluripotent stem cells by going through an MESP1 mesoderm progenitor stage. Methods: Mesp1 is a key transcription factor that regulates the development of early cardiovascular tissue. Using an MESP1-mTomato knock-in reporter human embryonic stem cell line, we compared the gene expression profiles of MESP1(+) and MESP1(-) cells and identified new signaling pathways that may promote endothelial differentiation. We also used a 3D scaffold to mimic the in vivo microenvironment to further improve the efficiency of endothelial cell generation. Finally, we performed cell transplantation into a critical limb ischemia mouse model to test the repairing potential of endothelial-primed MESP1(+) cells. Results: MESP1(+) mesoderm progenitors, but not MESP1(-) cells, have strong endothelial differentiation potential. Global gene expression analysis revealed that transcription factors essential for early endothelial differentiation were enriched in MESP1(+) cells. Interestingly, MESP1 cells highly expressed Sphingosine-1-phosphate (S1P) receptor and the addition of S1P significantly increased the endothelial differentiation efficiency. Upon seeding in a novel 3D microniche and priming with VEGF and bFGF, MESP1(+) cells markedly upregulated genes related to vessel development and regeneration. 3D microniches also enabled long-term endothelial differentiation and proliferation from MESP1+ cells with minimal medium supplements. Finally, we showed that transplanting a small number of endothelial-primed MESP1(+) cells in 3D microniches was sufficient to mediate rapid repair of a mouse model of critical limb ischemia. Conclusions: Our study demonstrates that combining MESP1(+) mesoderm progenitor cells with tissue-engineered 3D microniche and a chemically defined endothelial induction medium is a promising route to maximizing the production of endothelial cells in vitro and augment their regenerative power in vivo.
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