The upregulation of Gata transcription factors family and FOG-1 in expanded and differentiated cord blood-derived CD34+ hematopoietic stem cells to megakaryocyte lineage during co-culture with cord blood mesenchymal stem cells

Transfus Apher Sci. 2022 Dec;61(6):103481. doi: 10.1016/j.transci.2022.103481. Epub 2022 Jun 6.

Abstract

Umbilical cord blood (UCB) has improved into an attractive and alternative source of allogeneic hematopoietic stem cells (all-HSCs) in clinics and, research for three decades. Recently, it has been shown that the limited cell dose of, this valuable source can be enhanced by the ex vivo expansion of cells in many, ways. We evaluated the expression of the Gata transcription factors family and FOG-1, in expanded and differentiated cord blood-derived CD34 + hematopoietic stem cells to, megakaryocytes lineage., Methods: Separated mononuclear cells were cultured in DMEM complete medium., Harvested cells as a mesenchymal stem cell at 85 % confluency were cultured with, trypsin/EDTA and in 24-well plates. The characteristic analyses of isolated UCB- MSCs, were done by flow cytometry and adipogenic, chondrogenic, and osteogenic, differentiation assays. MACS purified UCB-CD34 + hematopoietic cells cultivated and, differentiated to megakaryocyte progenitor cells in the presence of cytokine cocktail, with UCB-MSCs. Then, the GATA1, GATA2, GATA3, and FOG-1 genes expression, after differentiation to megakaryocyte progenitor cells were performed by quantitative, real-time polymerase chain reaction (PCR)., Results: In this study, the results of real-time-PCR showed that the fold change, expression of GATA-1, FOG-1, and GATA-2 genes after co-culturing with UCB-MSCs, significantly increased to 7.3, 4.7, and 3.3-fold in comparison with control groups;respectively., Conclusion: UCB-MSCs can increase the expansion and differentiation of UCBCD34 + , to megakaryocyte progenitor cells through upregulation of GATA-1, GATA-2, and FOG-1 gene expression.

Keywords: Coculture; Cord blood stem cell; FOG-1; Gata transcription factor family; Mesenchymal stem cells; Upregulation.

MeSH terms

  • Antigens, CD34 / metabolism
  • Cell Differentiation
  • Cell Proliferation
  • Cells, Cultured
  • Coculture Techniques
  • Fetal Blood*
  • GATA Transcription Factors / metabolism
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Megakaryocytes / metabolism
  • Mesenchymal Stem Cells* / metabolism
  • Up-Regulation

Substances

  • Antigens, CD34
  • GATA Transcription Factors
  • ZFPM1 protein, human
  • GATA1 protein, human
  • GATA2 protein, human