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RESEARCH ARTICLE
Year : 2019  |  Volume : 4  |  Issue : 4  |  Page : 116-120

Isolation and culture of vascular wall-resident cd34+ stem/progenitor cells


1 Department of Physiology, School of Basic Medicine Science, Hubei University of Medicine; Hubei Key Laboratory of Embryonic Stem Cell Research, Hubei University of Medicine, Shiyan, Hubei, China
2 Department of Physiology, School of Basic Medicine Science, Hubei University of Medicine; Hubei Key Laboratory of Embryonic Stem Cell Research, Hubei University of Medicine; Department of Cardiology, Institute of Clinical Medicine, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, China

Correspondence Address:
Jun-Ming Tang
Department of Physiology, School of Basic Medicine Science, Hubei University of Medicine, Hubei, 442000, China, Hubei Key Laboratory of Embryonic Stem Cell Research, Hubei University of Medicine Shiyan, Hubei 442000; Department of Cardiology, Institute of Clinical Medicine, Renmin Hospital, Hubei University of Medicine, Hubei 442000
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/cp.cp_19_19

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Objective: The aim of this study is to observe the maintenance of stem cell properties of purified CD34-positive cells in vessel walls during in vitro expansion. Materials and Methods: Cells that migrated from the adventitial tissues of rat were collected and purified by microbead selection method to obtain CD34+ vascular wall-resident stem (VRS)/progenitor cell (PC). Those purified CD34+ VRS/PCs were evaluated by flow cytometry and immunofluorescent staining. The CD34+ VRS/PCs were continuously cultured until passage P3. Each passaged cell was evaluated by flow cytometry with anti-CD34. Results: After microbead selection, the CD34+ cells reached 88.07% ± 4.36% and these cells expressed neither endothelial (CD31) nor mature smooth muscle cell (smooth muscle-myosin heavy chain and SM22α) markers. Incubation of the purified CD34+ VRS/PCs at a density of 1.5 × 105 cells/100-mm dish, resulted in a gradual reduction of CD34-positive traits when passaged in vitro, starting at P1. Interestingly, the purified primary CD34+ VRS/PCs at a density of 1.0 × 104 cells per 100-mm dish show the traits of colony form growth, and P1 passaged cells were 79.2% ± 2.15% positive for CD34, then gradually lost the traits of CD34-positive cells when passaged in vitro. Conclusions: High purity CD34+ VRS/PCs can be obtained by magnetic bead screening. In vitro, low cell densities contribute to the maintenance of CD34+ VRS/PC traits.


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