Study on the optimal condition for culturing human stromal cells derived from umbilical cord blood
- VernacularTitle:人脐血源基质细胞分离培养条件的优化
- Author:
Cheng ZHANG
;
Xinghua CHEN
;
Lei GAO
- Publication Type:Journal Article
- Keywords:
fetal blood;
human umbilical cord blood-derived stromal cells;
cell separation;
cell culture
- From:
Medical Journal of Chinese People's Liberation Army
1982;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the optimal condition for culturing human umbilical cord blood-derived stromal cells(hUCBDSCs),and to observe their biological behaviors.Methods The umbilical cord blood was obtained from the Department of Obstetrics of the authors' hospital.The influence on the growth of hUCBDSCs was determined by the isolation method,the medium and the time of renewal of first medium were analyzed.The status of cell growth was observed under inverted microscope and the morphological characters were studied with the cells stained by Wright's staining.The hUCBDSCs were identified by cytochemistry and immunocytochemistry methods.Results The gelatin precipitation was better than other techniques for isolation.The optimal time for first medium renewal was the fourth day of culturing,and the improved Dexter-type cultural system was better than the classical Dexter-type cultural system in primary culture.The colonies of adherent cells began to form in 9-14 days(with a median of 12.1 days),and the number of colonies reached it maximum in 15-21 days(mean 19.4 days).On day 28,adherent cells spread all over the bottom of dish.On day 28 of culturing,these cells were found under light microscopy to have differentiated into three kinds of cells: fibroblast-like cells,macrophage-like cells and small-round cells.Cytochemistry assay revealed that the positive rate reached 100% with non-specific esterase(NSE) and saccharogen(PAS) staining.26% of the hUCBDSCs were positive with alkaline phosphatase(ALP) staining,but negative with peroxydase(POX) staining.Immunocytochemistry staining revealed that the positive rates of hUCBDSCs for CD31,CD68 and Fn were 96% and 95%,and 94% respectively,and for CD45 was 0%.Conclusion The hUCBDSCs could be successfully cultured in vitro,and it sets a foundation for further study on the clinical application of hUCBDSCs.
