Objective This study aimed to establish a standard method to derive and prepare feeder cells for the optimized growth of human embryonic stem cells(ESCs) and induced pluripotent stem cells(iPSCs). Methods CF-1 mouse fetuses between 12.5 and 14.5 days of gestation were separated, trypsinized, and cultured to derive primary mouse embryonic fibroblasts(MEFs). MEFs were then treated with mitomycin C to prepare feeder cells. The number of cells needed for high-quality feeder cells was determined through a variety of cell culture vessels. Results MEF from CF-1 pregnant mouse treated with mitomycin C could be used as feeder cells to support the culture and to maintain the undifferentiating of ESCs and iPSCs. High-quality feeder cells could be derived if an appropriate number of cells were plated. Conclusion MEF prepared in this method can optimally support and maintain the undifferentiated human ESCs and iPSCs. Furthermore, this method can be utilized as a standard method to derive feeder cells.
