Generation and Evaluation of Human Umbilical Cord Derived Mesenchymal Stem Cells with Antioxidant Capacity
10.19746/j.cnki.issn1009-2137.2024.06.039
- VernacularTitle:具有抗氧化能力的人脐带间充质干细胞制备及评价策略研究
- Author:
Xiao-Yu ZHANG
1
;
Pei-Lin LI
;
Jie TANG
;
Zhi-Ling LI
;
Rui-Cong HAO
;
Xiao-Tong LI
;
Wen-Jing ZHANG
;
Shi-Rong ZHAO
;
Li DING
;
Wen-Qing WU
;
Heng ZHU
Author Information
1. 河北大学生命科学院,河北保定071000;军事科学院军事医学研究院辐射医学研究所,北京 100850
- Keywords:
umbilical cord mesenchymal stem cells;
Wharton's jelly;
serum-free culture system;
collagenase-digested tissue debris culture protocol;
antioxidant
- From:
Journal of Experimental Hematology
2024;32(6):1888-1895
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare mesenchymal stem cells with antioxidant capacity (AO-MSC ) from human umbilical cords and evaluate its cell biological properties.Methods:In control group,mesenchymal stem cells (MSC) were isolated by digesting human umbilical cord Wharton's Jelly tissues with 0.2％ collagenase Ⅱ,and the released cells were collected and cultured in an animal serum-free culture medium.In AO-MSC group,incompletely collagenase Ⅱ-digested tissue debris were allowed to adhere to flusk flat bottoms and the AO-MSC was harvested by adherent culture. The conventional digestion and culture method was used as control.MSC colony forming ability was evaluated by fibroblast colony forming assay (CFU-F).MSC proliferative capacity was evaluated by CCK-8 assay.The MSC surface markers were detected by using flow cytometry and immunofluorescence staining.The adipogenic and osteogenic capacity of MSC was evaluated by multi-differentiation in vitro,and the mRNA expression of genes that control adipogenic and osteogenic differentiation was detected by real-time fluorescence quantitative PCR (RT-qPCR );Moreover,the mRNA expression of antioxidant substances such as SOD-1,GSH,GAT,and NQO1 in MSC was also evaluated by RT-qPCR.Results:The AO-MSC isolated by this strategy reached a confluence of 80％-90％ at around 18 days and grew in a swirling pattern.Flow cytometry and immunofluorescence staining assays showed that CD73,CD29,CD105,CD90 were highly expressed and CD31,CD45,HLA-DR were scarcely expressed in AO-MSC.AO-MSC exhibited stronger self-renewal and differentiation ability compared to MSC.However,the in vitro adipogenic-osteogenic capacity of MSC in the control group was stronger than that of AO-MSC.RT-qPCR assay showed that AO-MSC expressed higher mRNA levels of antioxidant substances compared to MSC.Conclusion:Human AO-MSC is successfully prepared from human umbilical cord without animal serum.
