In the culture system using human feeder cells, the mechanism through which these cells support undifferentiated growth of embryonic stem cells (ESCs) has not been well investigated. Here, we explored the mechanisms of 3 kinds of human feeder cells, including human placental cells from the chorionic plate, human bone marrow stromal cells, and human foreskin fibroblasts. First, we determined that undifferentiated growth of 2 kinds each of human (H1 and HSF6) and mouse (D3 and CE3) ESCs was possible in all human feeder cell types tested (human placental cells, human bone marrow stromal cells, and human foreskin fibroblasts), without the need for exogenous cytokine supplementation including basic fibroblast growth factor (bFGF) and leukemia inhibitory factor. We then prepared their corresponding endogenous bFGF-knockout feeders using siRNA and tried to maintain human and mouse ESCs in their undifferentiated state; however, neither human nor mouse ESCs could be maintained in bFGF-knockout human feeder cells. The expressions of stemness markers such as Oct-4 and Nanog were significantly decreased in the bFGF-knockout group compared with those in the controls, and differentiation had already occurred, despite the undifferentiated morphologic appearance of the ESCs. In conclusion, human feeder cells are able to support the undifferentiated growth of human and mouse ESCs via bFGF synthesis. Further, a bFGF-dependent pathway might be crucial for maintaining the undifferentiated characteristics of mouse and human ESCs.
The data presented herein support “Generation of an induced pluripotent stem cell line KUMCi001-A from CD34+ bone marrow cells of a patient with acute lymphoblastic leukemia using human placenta-derived cell conditioned medium.” The supplementary data were as follows. (1) Comparison of reprogramming efficiency of human placenta-derived cell conditioned medium with defined medium (mTeSR™1) and the generation of induced pluripotent stem cells (iPSCs) from a patient with acute lymphoblastic leukemia (ALL) with significantly higher reprogramming efficiency than that of the defined medium (
P
≤ 0.05). (2) Evaluation of differentiation capability of the generated ALL_iPSCs into hematopoietic stem cells (HSCs) and comparison with normal iPSCs using the colony-forming unit (CFU) assay. ALL_iPSCs manifested all lineages for hematopoiesis in their colonies similar to normal iPSCs. (3) ALL_iPSCs showed a considerably higher number of burst-forming unit-erythroid colonies indicating the presence of more erythroid progenitors than normal iPSCs; this tendency was confirmed in the CFU assay of ALL_CD34+ cells. This has been previously reported as a feature of ALL. Thus, the hematopoietic characteristics of the donor patient with ALL appear to be maintained in our ALL_hiPSC line although the karyotype was normalized during reprogramming.
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