Generation of induced pluripotent stem cells from peripheral blood cells using small molecules

Generation of induced pluripotent stem cells (iPSCs) using safe and non-viral methods play pivotal role in tissue engineering and regenerative medicine. In current study, we aimed to generate iPSCs from peripheral blood mononuclear cells (PBMC) in the presence of different small molecule and chemical compounds. PBMCs were isolated using Ficoll gradient method and their proliferation was induced by lipopolysaccharide (LPS). Reprogramming of extracted PBMCs was mediated using a combination of small molecule such as 5-Azacitidine, Kenpaullone, sodium butyrate and RepSox, which are generally epigenetic modulator of genes (OCT4, Sox2, Klf4, c-Myc) involved on pluripotent cell. Then, resulted pluripotent stem cells were cultured and identified on feeder MEFs. LPS effectively caused an increase in the number of PMBCs after 5 hours. Blood cells were successfully converted to iPSCs after treatment with different chemical compounds and small molecules for 14 days and then transferred on supportive mouse embryonic fibroblasts. The use of small molecules is a safe and cost-effective method for generating iPSCs from PBMCs. Mouse embryonic fibroblasts serve as a feeder layer for production of growth factors and preserving pluripotency of stem cells.