期刊
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
卷 59, 期 7, 页码 3028-3036出版社
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
DOI: 10.1167/iovs.17-23627
关键词
human embryonic stem cells; human corneal endothelial cells; differentiation; TGF beta-2; conditioned medium; neural crest
资金
- National Key R&D Program of China (Beijing, China) [2017YFA0103204]
PURPOSE. A shortage of human corneal endothelial cells (HCEC) for transplant and current methods of differentiation induction require chemical compounds, which might cast further influences after differentiation induction. Therefore, we developed a simple and straightforward approach to endothelial cell differentiation from human embryonic stem cells (hESC). METHODS. HESC are used to differentiate into HCEC by employing a two-stage method, which involves the application of two different types of conditioned culture medium, human corneal stromal cell-conditioned medium (HCSC-CM) and lens epithelial cell (LEC) plus HCSC-CM (LEC-CM+HCEC-CM). In brief, hESCs were treated with different conditioned media to induce directed endothelial differentiation. RESULTS. In the presence of conditioned culture medium, embryonic stem cells differentiate first under the control of periocular mesenchymal precursors (POMPs). Consequently, the expression of several POMP markers was observed. Following this first stage differentiation, POMPs were further directed to differentiate into corneal endothelial cell (CEC)-like cells in the presence of the second-conditioned culture medium. The differentiation of POMPs into CEC-like cells is regulated by a TGF beta-2/FOXC1 signaling pathway that is activated by the factors present in the conditioned culture medium. CONCLUSIONS. HCEC-like cells could be differentiated from hESC by simply using a two-step, preconditioned, medium-mediated approach, which could significantly minimize the workload to generate HCEC for potential clinical use. This research may provide an ideal cell source for corneal regenerative medicine and clinical treatment for corneal diseases in the future.
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