Int J Stem Cells.  2016 May;9(1):44-52. 10.15283/ijsc.2016.9.1.44.

Comparative Evaluation for Potential Differentiation of Endothelial Progenitor Cells and Mesenchymal Stem Cells into Endothelial-Like Cells

• Sabry D
• Noh O
• Samir M

• KMID: 2164160
• DOI: http://doi.org/10.15283/ijsc.2016.9.1.44

Abstract

Understanding the mechanisms of vascular remodeling could lead to more effective treatments for ischemic conditions. We aimed to compare between the abilities of both human Wharton jelly derived mesenchymal stem cells (hMSCs) and human cord blood endothelial progenitor cells (hEPCs) and CD34+ to induce angiogenesis in vitro. hMSCs, hEPCs, and CD34+ were isolated from human umbilical cord blood using microbead (MiniMacs). The cells characterization was assessed by flow cytometry following culture and real-time PCR for vascular endothelial growth factor receptor 2 (VEGFR2) and von Willebrand factor (vWF) to prove stem cells differentiation. The study revealed successful isolation of hEPCs, CD34+, and hMSCs. The hMSCs were identified by gaining CD29+ and CD44+ using FACS analysis. The hEPCs were identified by having CD133+, CD34+, and KDR. The potential ability of hEPCs and CD34+ to differentiate into endothelial-like cells was more than hMSCs. This finding was assessed morphologically in culture and by higher significant VEGFR2 and vWF genes expression (p<0.05) in differentiated hEPCs and CD34+ compared to differentiated hMSCs. hEPCs and CD34+ differentiation into endothelial-like cells were much better than that of hMSCs.

Keyword

Human endothelial progenitor cells; Human mesenchymal stem cells; CD34+; Endothelial cell differentiation

MeSH Terms

Fetal Blood
Flow Cytometry
Humans
Mesenchymal Stromal Cells*
Microspheres
Real-Time Polymerase Chain Reaction
Stem Cells*
Vascular Endothelial Growth Factor Receptor-2
von Willebrand Factor
Wharton Jelly
Vascular Endothelial Growth Factor Receptor-2
von Willebrand Factor

Figure

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  Fig. 1 (A) Undifferentiated EPCs cultured on fibronectin plate. After 14-day culture, a spindle-shaped (black arrow) and cobblestone-like (white arrow) morphology is typical for EPCs. (B) Spindle-shaped (black arrows) MSCs after two weeks in culture with 80~90% confluence. (C) CD34+ cells in culture at 0 day after isolation from hUC using microbeads. CD34+ cells were rounded nonadherent cells.

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  Fig. 2 FACS characterization of EPCs. Cells were uniformly positive for specific cell surface markers CD133, CD34, and KDR.

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  Fig. 3 FACS characterization of MSCs. Cells were uniformly negative for CD34 and CD45 and positive for CD44 and CD29.

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  Fig. 4 (A) The cluster formation (black arrows) and tubular network formation (white arrows) were highly identified in EPCs differentiated into endothelial-like cells. (B) The cluster formation (black arrows) and tubular network formation (white arrows) were identified in MSCs differentiated into endothelial-like cells. (C) CD34+ differentiated in culture after 14 days into endothelial-like cells. CD34+ differentiated as clusters (black arrows) and tubular network formation (white arrows).

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  Fig. 5 Quantitative RT-PCR of VEGFR2 gene expression in vitro in all studied groups. †Statistically significant compared to corresponding value in differentiated EPCs group (p<0.05). ‡Statistically significant compared to corresponding value in differentiated MSCs group (p<0.05).

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  Fig. 6 Quantitative RT-PCR of von Willebrand gene expression in vitro in all studied groups.

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  Fig. 7 FACS analysis of CD31 in all studied groups with high expression in differentiated EPCs and CD34+ compared to differentiated MSCs.

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  Fig. 8 FACS analysis of VE-cadherin in all studied groups with high expression in differentiated EPCs and CD34+ compared to differentiated MSCs.

Reference

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