Abstract
Chemically fixed mouse embryonic fibroblasts (MEFs), instead of live feeder cells, were applied to the maintenance of mouse induced pluripotent stem (miPS) cells. Formaldehyde and glutaraldehyde were used for chemical fixation. The chemically fixed MEF feeders maintained the pluripotency of miPS cells, as well as their undifferentiated state. Furthermore, the chemically fixed MEF feeders were reused several times without affecting their functions. These results indicate that chemical fixation can be applied to modify biological feeders chemically, without losing their original functions. Chemically fixed MEF feeders will be applicable to other stem cell cultures as a reusable extracellular matrix candidate that can be preserved on a long-term basis.
Highlights
Cultivation of pluripotent stem cells (PSCs) is inherently important for regenerative medicine
The results show that mouse induced pluripotent stem (miPS) cells cultured on GA-mouse embryonic fibroblasts (MEFs) or FA-fixed MEFs (FA-MEFs) exhibited similar levels of expression of Nanog–GFP (Fig. 3A); more than 95% of cells expressed Nanog–GFP, which suggests that miPS cells maintain their undifferentiated state on chemically fixed MEFs
The results show that miPS cells maintained their Alkaline phosphatase (AP) activity after six passages on chemically fixed MEF feeder cells (Fig. 3B)
Summary
Cultivation of pluripotent stem cells (PSCs) is inherently important for regenerative medicine. The undifferentiated state of cultured miPS cells was confirmed by Nanog expression and AP staining; the pluripotency of miPS cells was confirmed by neural differentiation induction and teratoma formation in mice.
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