Overexpressing NKx2.5 increases the differentiation of human umbilical cord drived mesenchymal stem cells into cardiomyocyte-like cells

Biomed Pharmacother. 2016 Mar:78:110-115. doi: 10.1016/j.biopha.2016.01.020. Epub 2016 Jan 24.

Abstract

Background: Nkx2.5 is one of the transcription factors in early myocardial cell development and Nkx2.5 gene expression increased gradually in the course of stem cells differentiation. In this study, this study aimed to investigate whether overexpression of NKx2.5 increases human umbilical cord drived mesenchymal stem cells (hUCMSCs) transdifferentiation into a cardiac phenotype in vitro.

Methods: hUCMSCs were transduced with Nkx2.5 at the third passage (transduced group). Gene expression of cTnI, Desmin, Nkx2.5 and GATA-4 in transduced group was analyzed using real-time PCR and immunohistochemistry, and compared with no-transduced hUCMSCs (control group), which were transfected with green fluorescent protein (GFP) only.

Results: Compared with control group, hUCMSCs in transduced group were shown by immunofluorescence to have higher expression of Nkx2.5. After incubation for 4 weeks, the mRNA and protein expression of cardiac genes, including cTnI, Desmin, Nkx2.5 and GATA-4, were up- regulated in transduced group compared with control group (P<0.05).

Conclusions: Overexpression of Nkx2.5 significantly promotes the differentiation of hUCMSCs into cardiomyocytes and increases the expression of cTnI, Desmin, Nkx2.5, and GATA-4.

Keywords: Cardiomyocyte-like cells; Human umbilical cord drived mesenchymal stem cells; Nkx2.5; Overexpression.

MeSH terms

  • Biomarkers / metabolism
  • Cell Differentiation* / genetics
  • Desmin / genetics
  • Desmin / metabolism
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • GATA4 Transcription Factor / genetics
  • GATA4 Transcription Factor / metabolism
  • Gene Expression Regulation
  • Homeobox Protein Nkx-2.5
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Humans
  • Infant, Newborn
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism*
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Real-Time Polymerase Chain Reaction
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transfection
  • Troponin I / genetics
  • Troponin I / metabolism
  • Umbilical Cord / cytology*

Substances

  • Biomarkers
  • Desmin
  • GATA4 Transcription Factor
  • Homeobox Protein Nkx-2.5
  • Homeodomain Proteins
  • NKX2-5 protein, human
  • Transcription Factors
  • Troponin I