Effects of Mesenchymal Stem Cells Feeder Layer, Culture Sera and Freeze-thaw Lysates on Expansion and Differentiation of Cord Blood CD34~+ Cells in Vitro
- VernacularTitle:间充质干细胞及其培养上清与融冻产物对脐血CD34~+细胞扩增及分化的作用
- Author:
Dunhua ZHOU
;
Ke HUANG
;
Yanfeng WU
;
Ting SHEN
;
Shaoliang HUANG
- Publication Type:Journal Article
- Keywords:
mesenchymal stem cells;
MSC culture sera;
MSC freeze-thaw lysates;
CD34~+ cells of cord blood
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2010;31(1):39-44
- CountryChina
- Language:Chinese
-
Abstract:
[Objective]To investigate the effects of mesenchymal stem cells (MSC) feeder layer, culture sere and freeze-thaw lysates on expansion and differentiation of cord blood CD34~+ cells in vitro. [Methods] MSC were isolated from human bone marrow and cultured until the third passage. Sera were obtained from the cultured MSC, and freeze-thaw lysates were obtained by repeated freeze-thaw procedures. Cord blood CD34~+ cells were isolated by magnetic cell separation system, and were co-cultured with the MSC feeder layer, culture sera, freeze-thaw lysates and hematopeietic growth factors (HGFs), respectively. The nucleated cells, CD34~+ cells, CD34~+CD38~- cells, CD41~+ cells and CD3~+ cells in the above culture system were detected by flow cytometry on day 6 and day 12. [Results] ①MSC feeder layer had a strong effect on nucleated cells, CD34~+,CD34~+CD38~- cells expansion. The MSC sera and freeze-thaw lysates had similar effect on cell expansion, but the effect was weaker than that of feeder layer (P<0.05). ② Both MSC sera and feeder layer inhibited cord blood CD34~+ cells differentiation toward CD3~+ cells or CD19~+ cells, and no significant differences were found between these two groups (P>0.05). ③ Both MSC sera and feeder layer promoted cord blood CD34~+ cells differentiation toward CD41~+ cells, and the effect was stronger in the feeder layer than that of the sera (P<0.05). ④ Freeze-thaw lysates had no effect on cell expansion and differentiation, and were similar with that of HGFs (P>0.05). [Conclusions] The MSC sera have positive effects on expansion of cord blood CD34~+ and CD34~+CD38~- cells, moreover they have the ability of promoting cord blood CD34~+ cells differentiation toward CD41~+ cells.
