The effect of decitabine on human induced pluripotent stem cells (hiPSCs) derived CD34+ cells expansion and the megakaryocytes generation and maturation.

Abstract

Background: Epigenetic modifiers play an important role in regulating the fate of hematopoietic stem cells (HSCs). The chromatin-modifying agents (CMA) have previously been shown to expand HSCs from cord blood (CB) and bone marrow (BM) CD34⁺ cells. Meanwhile, DNA methylation maintains persistent cellular memories and is thought to be the primary epigenetic barrier to reprogramming. The DNA hypomethylation drug decitabine is one of the CMA that could alter gene expression and HSC self-renewal. It has been reported that decitabine could promote platelets generation in ITP patients.

Objective: It's unknown if decitabine could affect CD34⁺ cells and megakaryocytes generation and maturation from human induced pluripotent stem cells (hiPSCs).

Methods: We utilized serum free, exon free and feeder free differentiation system to generate CD34⁺ from hiPSCs and induced them differentiation into megakaryocytes. Different concentrations of decitabine were added at different stages and analyzed these cells by RT-PCR, flow cytometry analysis, cell counting and other regular experimental methods.

Results: The proliferation and function of CD34⁺ cells in vitro were significantly suspended after exposure to decitabine. Low concentration of decitabine could maintain the CD34⁺ function. In addition, we found that decitabine did not have any effect on the megakaryocyte generation, but it prevented megakaryocyte maturation. The DNA methyltransferases (DNMTs) changed a lot not only in CD34⁺ stage but also in the megakaryocyte generation and maturation due to decitabine addition.

Conclusions: These results suggested that the effect of decitabine on CD34⁺ cells from hiPSCs was very different from CB, PB and BM CD34⁺ cells and the epigenetic changes may play an important role in the CD34⁺ expansion and megakaryocytes maturation. It may provide a potential mechanism of studying hiPSCs derived HSCs and megakaryocytes maturation in the future.