Induction Of Pluripotent Stem Cells From Cancer Patient Adipose Stem Cells By Defined Factors

Induced pluripotent stem cells (iPS cells) are derived from somatic cells that regain pluripotency by the nuclear programming with exogenous factors. iPS cells have immense potential applications in establishing disease models and understanding disease mechanisms, drug screening and assessments, cell therapies, etc. avoiding both immunologic and ethical barriers to their use. So far, rats, human, pigs, monkeys and rabbits'iPS cell lines are established; and confirmed that iPS cells can attain developmental pluripotency that is similar to that of ES cells; iPS–based disease models, drug discovery and assessments have made great achievements. However, it is necessary to overcome challenges such as low reprogramming efficiency and risk due to tumorigenicity, iPS–based cancer model hasn't been established.Adipose tissue is easy to obtain in large amounts with minimal morbidity, isolating adipose-derived stem cells is relatively simple and fast, besides they are easy to maintain in culture. Adipose-derived stem cells are a heterogeneous group of cell populations, which have a large potential for selfrenewal, while also maintaining the ability to differentiate a limited repertoire of cell types such as adipocytes, myocytes, osteoblasts and chondrocytes. We hypothesize that adipose-derived stem cells represent an intermediate state between differentiated and pluripotent cells, the state may be facilitate generation of human iPS cells.Here, our experiments'main methods as follows:(1)Plasmid(pMXs-Oct4,pMXs-Sox2,pMXs-Klf4,pMXs-c-Myc,pMXs-EGFP) DNA purification and pMX-based retrovirus production,(2)Adipose tissue acquisition, adipose-derived stem cells isolation and primary culture and identification,(3) pMX-based retrovirus transduced adipose-derived stem cells and culture of iPS Cells,(4) Culture and identification of adipose-derived iPS Cells (Alkaline Phosphatase Staining, Quantitative-PCR and Immunofluorescence Staining, etc.).Results: we reported that adult human adipose-derived stem cells from four 40 to 50-year-old individuals can be reprogrammed into induced pluripotent stem (iPS) cells with ectopic expression Oct4,Sox2,Klf4 and c-Myc. We found that adipose-derived stem cells can be efficiently reprogrammed into iPS cells, and can be effectively serve as feeder layers of their own iPS cells. Generation of iPS lines from adipose-derived stem cells may be advantageous in providing platforms for iPS cells technology and diease models for cancer.