Establishment Of A Culture System For Human Induced Pluripotent Stem Cells (iPS Cells) On Microfluidic Chip

Induced pluripotent stem cells (iPS cells) were derived from somatic cells by introducing specific transcription factors, which made somatic cells reprogrammed. The emergence of iPS cells was a revolution in life science. Studying iPS cells on microfluidic chip which has the advantages of miniaturization, integration and portabilitycan promotes the development of iPS cells greatly.The aims of our study are establishing a stable culture system of human induced pluripotent stem cells on microfluidic chip and studying how the pH solution influence the FDA entering cells on microfluidic chip preliminarily.The main work of our research is as follows:1. We isolated and cultured the mouse embryonic fibroblast, and obtained the feeder layer suitable for the growth of human induced pluripotent stem cells by treating the third passage of mouse embryonic fibroblasts with10μ g/mL mitomycin C for2.5hours.2. Exploring the characteristics of human induced pluripotent stem cells clone, we established a culture system of human induced pluripotent stem cells and successfully recovered the cell after cryopreserved in vitro.3. We designed and fabricated a glass-glass microfluidic chip and a PDMS-glass microfluidic chip, and cultured mouse embryonic fibroblast and human induced pluripotent stem cells on the chip, which provided a basis for further study of stem cells differentiation on chips.4. No matter in petri dishes or on microfluidic chip, undifferentiated human induced pluripotent stem cells have the positive alkaline phosphatase (AKP) staining, which illustrates that establishing a stable culture system of iPS cells on microfluidic chip is feasible.5. Studying the changes on FDA entering cells with different pH (pH4, pH5and pH6) and different time (10min,20min and30min) treatment suggests that pH may change the cell membrane permeability or esterase activity.