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Isolation, characterisation and comparative analysis of human umbilical cord vein perivascular cells and cord blood mesenchymal stem cells

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Abstract

Perivascular cells are known to be ancestors of mesenchymal stem cells (MSCs) and can be obtained from heart, skin, bone marrow, eye, placenta and umbilical cord (UC). However detailed characterization of perivascular cells around the human UC vein and comparative analysis of them with MSCs haven’t been done yet. In this study, our aim is to isolate perivascular cells from human UC vein and characterize them versus UC blood MSCs (UCB-MSCs). For this purpose, perivascular cells around the UC vein were isolated enzymatically and then purified with magnetic activated cell sorting (MACS) method using CD146 Microbead Kit respectively. MSCs were isolated from UCB by Ficoll density gradient solution. Perivascular cells and UCB-MSCs were characterized by osteogenic and adipogenic differentiation procedures, flow cytometric analysis [CD146, CD105, CD31, CD34, CD45 and alpha-smooth muscle actin (α-SMA)], and immunofluorescent staining (MAP1B and Tenascin C). Alizarin red and Oil red O staining results showed that perivascular cells and MSCs had osteogenic and adipogenic differentiation capacity. However, osteogenic differentiation capacity of perivascular cells were found to be less than UCB-MSCs. According to flow cytometric analysis, CD146 expression of perivascular cells were appeared to be 4.8-fold higher than UCB-MSCs. Expression of α-SMA, MAP1B and Tenascin-C from perivascular cells was determined by flow cytometry analysis and immunfluorescent staining. The results appear to support the fact that perivascular cells are the ancestors of MSCs in vascular area. They may be used as alternative cells to MSCs in the field of vascular tissue engineering.

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Acknowledgments

The authors wish to thank The Scientific and Technological Research Council of Turkey (Project Number: 113S815) for their financial support.

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Authors and Affiliations

  1. Department of Stem Cell Sciences, Graduate School of Health Sciences, Hacettepe University, 06100, Ankara, Turkey

    Beyza Gökçinar-Yagci & Betül Çelebi-Saltik

  2. Center for Stem Cell Research and Development, Hacettepe University, 06100, Ankara, Turkey

    Beyza Gökçinar-Yagci & Betül Çelebi-Saltik

  3. Department of Obstetrics and Gynecology, Hacettepe University Faculty of Medicine, 06100, Ankara, Turkey

    Özgür Özyüncü

Authors
  1. Beyza Gökçinar-Yagci
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  2. Özgür Özyüncü
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  3. Betül Çelebi-Saltik
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Corresponding author

Correspondence to Betül Çelebi-Saltik.

Ethics declarations

In this study, UC and UCB samples were obtained after birth of healthy babies borned in Hacettepe University Faculty of Medicine, Obstetrics and Gynecology Clinic. For the use of these non-invasive samples, permission was taken from Non-interventional Clinical Researches Ethics Committee.

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The authors declare that they have no conflict of interest.

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Gökçinar-Yagci, B., Özyüncü, Ö. & Çelebi-Saltik, B. Isolation, characterisation and comparative analysis of human umbilical cord vein perivascular cells and cord blood mesenchymal stem cells. Cell Tissue Bank 17, 345–352 (2016). https://doi.org/10.1007/s10561-015-9542-5

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  • DOI: https://doi.org/10.1007/s10561-015-9542-5

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