Isolation and characterization of human umbilical cord‑derived endothelial colony‑forming cells

Zhang,Hao; Tao,Yanling; Ren,Saisai; Liu,Haihui; Zhou,Hui; Hu,Jiangwei; Tang,Yongyong; Zhang,Bin; Chen,Hu

doi:10.3892/etm.2017.5035

Isolation and characterization of human umbilical cord‑derived endothelial colony‑forming cells

Authors:
- Hao Zhang
- Yanling Tao
- Saisai Ren
- Haihui Liu
- Hui Zhou
- Jiangwei Hu
- Yongyong Tang
- Bin Zhang
- Hu Chen
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Affiliations: Department of Hematopoietic Stem Cell Transplantation, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, P.R. China, Department of Hematology, Affiliated Hospital of Jining Medical University, Jining, Shandong 272000, P.R. China, Department of Graduate School, Jining Medical University, Jining, Shandong 272000, P.R. China
Published online on: August 25, 2017 https://doi.org/10.3892/etm.2017.5035
Pages: 4160-4166
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Endothelial colony‑forming cells (ECFCs) are a population of endothelial progenitor cells (EPCs) that display robust proliferative potential and vessel‑forming capability. Previous studies have demonstrated that a limited number of ECFCs may be obtained from adult bone marrow, peripheral blood and umbilical cord (UC) blood. The present study describes an effective method for isolating ECFCs from human UC. The ECFCs derived from human UC displayed the full properties of EPCs. Analysis of the growth kinetics, cell cycle and colony‑forming ability of the isolated human UC‑ECFCs indicated that the cells demonstrated properties of stem cells, including relative stability and rapid proliferation in vitro. Gene expression of Fms related tyrosine kinase 1, kinase insert domain receptor, vascular endothelial cadherin, cluster of differentiation (CD)31, CD34, epidermal growth factor homology domains‑2, von Willebrand factor and endothelial nitric oxide synthase was assessed by reverse transcription‑polymerase chain reaction. The cells were positive for CD34, CD31, CD73, CD105 and vascular endothelial growth factor receptor‑2, and negative for CD45, CD90 and human leukocyte antigen‑antigen D related protein according to flow cytometry. 1,1'‑dioctadecyl‑3,3,3',3'‑tetra‑methyl‑indocarbocyanine perchlorate‑labeled acetylated low‑density lipoprotein and fluorescein isothiocyanate‑Ulex europaeus‑l were used to verify the identity of the UC‑ECFCs. Matrigel was used to investigate tube formation capability. The results demonstrated that the reported technique is a valuable method for isolating human UC‑ECFCs, which have potential for use in vascular regeneration.

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