The effect of ADSC on the activity of EPC under co-culture and its related mechanism
10.19405/j.cnki.issn1000-1492.2023.04.005
- Author:
Le Cao
1
;
Zhenfei Ding
2
;
Kai Sun
1
;
Haitao Fan
1
;
Haitao Yang
1
Author Information
1. Dept of Orthopaedic，Fuyang Hospital Affiliated to Anhui Medical University，Fuyang 236000
2. Anhui Medical University Graduate School，Hefei 230032
- Publication Type:Journal Article
- Keywords:
adipose-derived stem cells;
endothelial progenitor cells;
PI3K / AKT
- From:
Acta Universitatis Medicinalis Anhui
2023;58(4):547-553
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of co-culture of adipose-derived stem cells(ADSC) and endothelial progenitor cells(EPC) on the activity of EPC and its related mechanism．
Methods:Rat ADSC and EPC were isolated，cultured，expanded and identified in vitro ．The experiment was divided into three groups : EPC group，EPC + ADSC co-culture group，and EPC + ADSC + PI3K-inhibitor group．After 48 hours of co-culture，the cells of the three groups were treated with Transwell．The effects of ADSC and EPC co-culture and PI3K / AKT pathway on EPC activity were evaluated by CCK-8 assay，scratch assay and angiogenesis assay，respectively．Western blot was used to detect vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS) ，vascular endothelial-cadherin ( VE-cadherin) ，CD133 ，phospho-phosphatidylinositol 3-kinase ( phospho-phosphatidylinositol 3-kinase(p-PI3K) and phospho-protein Kinase B (p-AKT) expression levels in EPC to detect the effects of ADSC and EPC co-culture and PI3K / AKT pathway on the differentiation ability of EPC into mature endothelial cells.
Results:CCK-8 results showed that the absorbance at 450 nm of EPC in EPC + ADSC co-culture group was higher than that in EPC group and EPC + ADSC + PI3K-inhibitor group at different time points，and the difference was statistically significant (P＜0. 01) ．The scratch test showed that the relative scratch distance of EPC + ADSC co-culture group was smaller than that of EPC group and EPC + ADSC + PI3K-inhibitor group after 24 hours ，and the difference was statistically significant (P＜0. 01) ．Tube formation assay showed that the average number of tubelike structures formed in EPC + ADSC co-culture group was higher than that in EPC group and EPC + ADSC + PI3K-inhibitor group，and the difference was statistically significant (P＜0. 01) ．Western blot showed that the expression levels of VEGFA，eNOS，VE-cadherin，p-PI3K and p-AKT of EPC in EPC + ADSC co-culture group were higher than those in EPC group and EPC + ADSC + PI3K-inhibitor group．The expression level of CD133 in EPC group was lower than that in EPC + ADSC + PI3K-inhibitor group，and the difference was statistically significant (P ＜0. 01) ．
Conclusion :Co-culture of ADSC and EPC can improve the proliferation，migration，differentiation and vasogenic activity of EPC through the regulation of PI3K / AKT pathway.
- Full text:2024071710113447452共培养下脂肪源性干细胞对内...细胞活性的影响及其相关机制_曹乐.pdf
