Construction and a preliminary study of paracrine effect of bone marrow-derived endothelial progenitor cell sheet

被引:3
作者
Xue, Fenlong [1 ]
Bai, Yunpeng [2 ]
Jiang, Yiyao [1 ,3 ]
Liu, Jianshi [4 ]
Jian, Kaitao [2 ,4 ]
机构
[1] Tianjin First Cent Hosp, Dept Cardiovasc Surg, Tianjin 300192, Peoples R China
[2] Tianjin Chest Hosp, Dept Cardiovasc Surg, Tianjin 300051, Peoples R China
[3] Bengbu Med Coll, Affiliated Hosp 1, Dept Cardiovasc Surg, Bengbu 233004, Anhui, Peoples R China
[4] DeltaHlth Hosp Shanghai, Dept Cardiovasc Surg, Shanghai 200336, Peoples R China
基金
中国国家自然科学基金;
关键词
Bone marrow; Cell sheet; Endothelial progenitor cell; Paracrine action; SDF-1; alpha/CXCR4; axis; SMOOTH-MUSCLE-CELL; TRANSPLANTATION; TECHNOLOGY; EXPRESSION; HEART; FABRICATION; MIGRATION; PATHWAY; TISSUES; REPAIR;
D O I
10.1007/s10561-021-09932-w
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The release of paracrine factors from endothelial progenitor cell (EPC) sheet is a central mechanism of tissue repair. The purpose of this study was to constuct the rat bone marrow derived-endothelial progenitor cell (BM-EPCs) sheet and investigate invest the role of stromal cell-derived factor-1 alpha (SDF-1 alpha)/CXCR4 axis in the biological function of BM-EPCs sheet. BM-EPC cells were identified by the cell-surface markers-CD34/CD133/VE-cadherin/KDR using flow cytometry and dual affinity for acLDL and UEA-1. After 7 days of incubation, the BM-EPC single-cell suspensions were seeded on thermo-sensitive plate to harvest the BM-EPC cell sheets. The expression levels of SDF-1 alpha/CXCR4 axis-associated genes and proteins were examined using RT-qPCR and western blot analysis, and enzyme-linked immunosorbent assay (ELISA) was applied to determine the concentration of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and SDF-1 alpha in the cell culture medium. The BM-EPC cell sheets were successfully harvested. Moreover, BM-EPC cell sheets have superior migration and tube formation activity when compared with single cell suspension. When capillary-like tube were formed from EPCs sheets, the releasing of paracrine factors such as VEGF, EGF and SDF-1 alpha were increased. To reveal the mechanism of tube formation of BM-EPCs sheets, our research showed that the activation of PI3K/AKT/eNOS pathway was involved in the process, because the phosphorylation of CXCR, PI3K, AKT and eNOS were increased. BM-EPC cell sheets have superior paracrine and tube formation activity than the BM-EPC single-cell. The strong ability to secrete paracrine factors was be potentially related to the SDF-1 alpha/CXCR4 axis through PI3K/AKT/eNOS pathway.
引用
收藏
页码:185 / 197
页数:13
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