Feeder-Free Culture of Murine Induced Pluripotent Stem Cells

Induced pluripotent stem cells (iPSCs), reprogrammed from somatic cells, can self-renew and differentiate into all cell types of the adult body. Typically, murine iPSCs are co-cultured with feeder cells, which supply various undefined growth factors and extracellular matrix components. To ensure experimental results stem exclusively from iPSCs, we aimed to establish a feeder-independent culture system suitable for maintaining murine fibroblast-derived iPSCs. Here, we have investigated culturing murine iPSCs on gelatin-coated plates in the presence of leukemia inhibitory factor (LIF). After five passages, most iPSCs that were previously established and maintained on feeder cells adapted to the feeder-cell-free culture conditions. Morphological characteristics of iPSCs cultured in the presence or absence of feeder cells were similar, with compact colonies and cytoplasmic membrane contact among the cells. Additionally, the absence of feeder cells had no impact on the proliferation of iPSCs. Importantly, the expression of pluripotency and differentiation markers in murine iPSCs was not affected when cultured in the absence of feeder cells after thirty passages. These results suggest that iPSCs cultured without feeder cells maintain their pluripotency. In summary, we have successfully developed a highly efficient system for cultivating murine iPSCs without feeder cells.