Differentiation of Adipose Tissue–Derived CD34+/CD31− Cells into Endothelial Cells In Vitro

  • Anoosha Forghani
  • Srinivas V. Koduru
  • Cong Chen
  • Ashley N. Leberfinger
  • Dino J. Ravnic
  • Daniel J. HayesEmail author


In this study, CD34+/CD31 progenitor cells were isolated from the stromal vascular fraction (SVF) of adipose tissue using magnetic activated cell sorting. The endothelial differentiation capability of these cells in vitro was evaluated by culturing them in vascular endothelial growth factor (VEGF)–induced medium for 14 days. Viability, proliferation, differentiation, and tube formation of these cells were evaluated. Cell viability study revealed that both undifferentiated and endothelial-differentiated cells remained healthy for 14 days. However, the proliferation rate was higher in undifferentiated cells compared to endothelial-differentiated ones. Upregulation of endothelial characteristic genes (von Willebrand factor (vWF) and VE-cadherin) was observed in 2D culture. However, PECAM (CD31) was only found to be upregulated after the cells had formed tube-like structures in 3D Matrigel culture. These results indicate that adipose-derived CD34+/CD31 cells when cultured in VEGF-induced medium, are capable differentiation into endothelial-like lineages. Tube formation of the cells started 3 h after seeding the cells on Matrigel and formed more stable and connected network 24 h post-seeding in presence of VEGF.

Lay Summary

In this work, endothelial differentiation capability of CD34+/CD31 cells isolated from adipose tissue was evaluated. The results showed that these cells can be successfully differentiated into endothelial cells in vitro in the presence of VEGF. Vascularization is crucial for the survival of the cells and formation of new tissues in engineered tissue constructs. Therefore, finding a proper source of progenitor cells capable of endothelial differentiation is of substantial importance in tissue engineering for the vascularization of large-volume complex tissues. CD34+/CD31 cells can be easily harvested from adipose tissues in abundance unlike other mesenchymal stem cells such as bone marrow stem cells, which makes them advantageous for vascularization in tissue engineering.


Stromal vascular fraction CD34+ cells Endothelial differentiation Adipose tissue 


Funding Information

Research reported in this article was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health under award number (RDE024790A).

Compliance with Ethical Standards


The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Supplementary material

40883_2019_93_MOESM1_ESM.docx (551 kb)
ESM 1 (DOCX 550 kb)


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Copyright information

© The Regenerative Engineering Society 2019

Authors and Affiliations

  • Anoosha Forghani
    • 1
  • Srinivas V. Koduru
    • 2
  • Cong Chen
    • 1
  • Ashley N. Leberfinger
    • 2
  • Dino J. Ravnic
    • 2
  • Daniel J. Hayes
    • 1
    • 3
    • 4
    Email author
  1. 1.Department of Biomedical Engineering, Millennium Science ComplexPennsylvania State UniversityUniversity ParkUSA
  2. 2.Department of Surgery, College of MedicinePenn State Health Milton S. Hershey Medical CenterHersheyUSA
  3. 3.Materials Research Institute, Materials Characterization Lab, Millennium Science ComplexPennsylvania State UniversityUniversity ParkUSA
  4. 4.The Huck Institute of the Life Sciences, Millennium Science ComplexPennsylvania State UniversityUniversity ParkUSA

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