Expansion and identification of primary rat aortic vascular stem cells in vitro
10.12122/j.issn.1673-4254.2024.10.06
- VernacularTitle:原代大鼠主动脉血管干细胞的体外培养扩增及鉴定
- Author:
Huagen MA
1
,
2
;
Yan HUANG
;
Yingxin YANG
;
Haiqin LIU
;
Yuanyu TANG
;
Weihong CONG
Author Information
1. 中国中医科学院西苑医院,北京 100091
2. 北京中医药大学中医学院,北京 100029
- Keywords:
vascular stem cells;
aorta;
adherent culture of the segmented outer membrane;
primary culture;
immunophenotype;
differentiation potential;
vascular remodeling
- From:
Journal of Southern Medical University
2024;44(10):1881-1886
- CountryChina
- Language:Chinese
-
Abstract:
Objective To culture and expand primary rat aortic vascular stem cells in vitro and evaluate their characteristics as mesenchymal stem cells.Methods The thoracic and abdominal aortas isolated from 2-to 3-week-old Sprague-Dawley rats were cut into vascular segments 2.0 mm in length and cultured in culture flasks till adhesion and solidification of the outer membranes.The primary cells were further cultured to 80%-90%confluence before passaging.The morphology and growth characteristics of the cells were observed under a microscope,and the expressions of surface marker CD molecules on the cells was analyzed using flow cytometry.Adipogenic and osteogenic differentiation assays were performed to assess the capacity of the cells for multilineage differentiation.Results After 3 days of culture,a small number of spindle,star-shaped or polygonal cells migrated out from the peripheral of the vascular segments.At 5-6 days,island-like cell clusters occurred and the cells began to proliferate rapidly.The cell clusters expanded radially and showed signs of cell cloning.At 7-8 days,the cells fused into sheets and displayed a vortex-like distribution.The cells in the third passage presented with a uniform morphology,showing a typical fibroblast-like arrangement.Flow cytometry showed that the cells expressed predominantly CD44(80.3%),CD73(62.2%)and CD90(46.8%)with low expressions of CD34(1.1%),CD45(0.2%)and CD11b/c(0.2%).Adipogenic and osteogenic differentiation experiments demonstrated that the cells were capable of lipogenic and osteogenic differentiation in vitro.Conclusion Rat aortic vascular stem cells with mesenchymal stem cell characteristics can be successfully isolated and cultured by adherent culture of the segmented outer membrane.
