Reprogramming of human peripheral blood mononuclear cells into induced mesenchymal stromal cells using non-integrating vectors

Mesenchymal stromal cells (MSCs) have great value in cell therapies. The MSC therapies have many challenges due to its inconsistent potency and limited quantity. Here, we report a strategy to generate induced MSCs (iMSCs) by directly reprogramming human peripheral blood mononuclear cells (PBMCs) with OCT4, SOX9, MYC, KLF4, and BCL-XL using a nonintegrating episomal vector system. While OCT4 was not required to reprogram PBMCs into iMSCs, omission of OCT4 significantly impaired iMSC functionality. The omission of OCT4 resulted in significantly downregulating MSC lineage specific and mesoderm-regulating genes, including SRPX, COL5A1, SOX4, SALL4, TWIST1. When reprogramming PBMCs in the absence of OCT4, 67 genes were significantly hypermethylated with reduced transcriptional expression. These data indicate that transient expression of OCT4 may serve as a universal reprogramming factor by increasing chromatin accessibility and promoting demethylation. Our findings represent an approach to produce functional MSCs, and aid in identifying putative function associated MSC markers. Induced MSCs (iMSCs) are generated by direct reprogramming of human PBMCs using 5 factor transfection, including OCT4, SOX9, MYC, KLF4, and BCL-XL.

Automated summary

Researchers have developed a method to generate induced mesenchymal stromal cells (iMSCs) by directly reprogramming human peripheral blood mononuclear cells (PBMCs) using five factors. Omission of OCT4, one of the factors, significantly impairs the functionality of iMSCs and downregulates MSC-specific and mesoderm-regulating genes. The transient expression of OCT4 increases chromatin accessibility and promotes demethylation.