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

Abstract

Introduction: 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.
Materials & methods: 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.
Results: 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.
Conclusion: 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.