İnsan fibroblast hücre hatlarından uyarılmış pluripotent kök hücre eldesi

Objective: Induced pluripotent stem cells (iPSCs) possess the remarkable abilities to differentiate into cell types from all three germ layers and to self-renew indefinitely. These features make them highly valuable for in vitro modeling of human biology and gene and cell therapies. However, current methods for iPSC generation are often inefficient and require prolonged culture periods, necessitating improved protocols with enhanced safety and efficacy. In this study, we aimed to reprogram HFF-1 human fibroblasts into iPSCs using integration-deficient lentiviral vectors (IDLV-OSKs) encoding the reprogramming factors Oct4, Sox2, and Klf4, along with sodium butyrate as a supporting small molecule to increase efficiency. Methods: In our study, IDLV-OSK vectors encoding the transcriptional regulators Oct4, Sox2, and Klf4 were used to reprogram HFF-1 fibroblast cells into pluripotency. In addition to the viral vector, the epigenetic modulator sodium butyrate, which is a histone deacetylase inhibitor, was used as a supportive molecule. With our applied reprogramming protocol, colonies suitable for passaging were obtained within 9–12 days. The resulting iPSCs were characterized morphologically and also by the expression of pluripotency markers expressed on the cell surface (alkaline phosphatase, Tra-1-81, Tra-1-60) and those located in the nucleus (Oct4, Klf4, c-Myc, Nanog). Results: We generated iPSCs using IDLV-OSKs with the support of a histone deacetylase inhibitor. The resulting colonies exhibited the expected morphological characteristics. The expression of surface and nuclear pluripotency markers was demonstrated in the colonies. Conclusion: Our findings demonstrate that IDLV-OSK vectors, along with sodium butyrate, constitute an efficient and potentially safe approach for reprogramming human fibroblasts into iPSCs with our protocol. This method holds promise for efficient and safer applications in disease modeling and the development of experimental therapeutic strategies. Keywords: Induced pluripotent stem cells, Sodium butyrate, Lentiviral vectors