Generation of Human-Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells using Small-Molecule Compound VC6TFZ

Abstract

BACKGROUND: Induced pluripotent stem cells (iPSCs) were generated from somatic cells through reprogramming process. Peripheral blood mononuclear cells (PBMNCs) were an attractive source cells due to the ease of accessibility, need minimal invasive procedure, and can be stored frozen. Small-molecule compound VC6TFZ has been successfully reprogrammed iPSCs from mouse fibroblast, but it has not been proven in human.
 AIM: This study aims to determine whether the small-molecule compound VC6TFZ can induce pluripotency of PBMNC to generate iPSCs.
 METHODS: Mononuclear cells were isolated from peripheral venous blood using centrifugation gradient density method. Mononuclear cells were cultured for 6 days in expansion medium and 48 h using hanging drop method. Pluripotency induction process using small-molecule compound VC6TFZ was done in 14 days then the medium changed to 2i medium for 7 days. Identification of iPSCs based on colony morphology and expression of pluripotency marker OCT4 and SOX2.
 RESULTS: Colonies appeared on day 9 of reprogramming process. These colonies had round, large, and cobble stone morphology like embryonic stem cell. These colonies had positive expression of pluripotency markers OCT4 and SOX2. All experimental groups had significantly higher expression of OCT4 and SOX2 than control group.
 CONCLUSION: Small-molecule compound VC6TFZ could induce pluripotency of human PBMNC to generate iPSCs.