Human embryonic stem cell-derived mesenchymal stem cells improved premature ovarian failure

Figure 1 Derivation and identification of human embryonic stem cell-derived mesenchymal stem cells and bone marrow-derived mesenchymal stem cells. A: Schematic presentation of the procedure used to derive human mesenchymal stem cells (MSCs) from embryonic stem (ES) cells. Colonies of ES cells were enzymatically detached and cultured for 10 d in suspension to form embryoid bodies, which were then plated onto gelatin-coated tissue culture plates. After 10 d, outgrowths of the cells that sprouted from embryoid bodies were mechanically isolated by a cell scraper and subsequently expanded in mesenchymal stem cell culture medium; B and C: Morphology and karyotype of ES-MSCs and BM-MSCs. Passage-5 ES-MSCs and BM-MSCs showed a fibroblastic morphology and normal karyotype; D: Alizarin red staining after 14 d of culture in osteogenic medium indicated the osteogenic differentiation potential of ES-MSCs and BM-MSCs (P4). Oil red staining after 21 d of culture in adipogenic medium showed the adipogenic differentiation potential of ES-MSCs and BM-MSCs (P4). Alcian blue staining after 21 d of culture in chondrogenic medium showed chondrogenic differentiation potential of ES-MSCs and BM-MSCs; E, F: Flow cytometric analysis indicated that cultured ES-MSCs and BM-MSCs expressed CD44, CD90, CD73 and endoglin (CD105), but not hematopoietic lineage markers CD11b, CD34 and protein tyrosine phosphatase receptor type C (CD45); G: ES-MSCs proliferated more rapidly than BM-MSCs. Results are expressed as mean ± standard error, ^aP < 0.05, ^bP < 0.01; n = 3-5. ES-MSCs: Embryonic stem cell-derived mesenchymal stem cells; BM-MSCs: Bone marrow-derived mesenchymal stem cells; EBs: Embryoid bodies; bFGF: Basic fibroblast growth factor; P: Passage.