Improved primary culture system increases the yield of human umbilical cord mesenchymal stem cells
10.3969/j.issn.2095-4344.2015.10.001
- VernacularTitle:改良原代培养体系提高人脐带间充质干细胞的产量
- Author:
Yanlin GAO
;
Ningkun ZHANG
;
Houliang CHEN
;
Lianru GAO
;
Zhiming ZHU
- Publication Type:Journal Article
- Keywords:
Umbilical Cord;
Mesenchymal Stem Cels;
Cels,Cultured
- From:
Chinese Journal of Tissue Engineering Research
2015;(10):1477-1481
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:At present, a lot of research about culture methods for umbilical cord mesenchymal stem cels, but not for the waste of primary system. OBJECTIVE:To explore the best culture method of human umbilical cord mesenchymal stem celsin vitro. METHODS:Human umbilical cord mesenchymal stem cels were prepared by tissue explants method, recorded as initial culture group. The centrifugal fluid and tissue of the primary culture flask were centrifuged and divided into three groups for secondary culture: tissue group, mixed group and pure liquid group. Cel morphology, time for cel acquisition, and yield of primary cels in the four groups were observed; the cel growth curve was analyzed by MTT assay; and cel cycle and phenotype were detected by flow cytometry. RESULTS AND CONCLUSION: The average time for cel acquisition in the initial culture group, tissue group, mixed group and pure liquid group were (15.00±0.45), (7.0±0.3), (8.00±0.25) and (8.00±0.25) days, respectively. The number of cels at first generation was (4.0±0.5)×105, (9.0±0.55)×105, (15.0±0.2)×105 and (7.0±0.33)×105 markers of the four groups had no significant differences. The human umbilical cord mesenchymal stem cels can be obtained rapidly and largely through the secondary culture to the primary culture system. T75 culture bottle, respectively. Under the inverted microscope, cels in the four groups were fusiform-like adherent cels, which were in paralel or circinate arrangement. Growth curve, proliferative activity, surface markers of the four groups had no significant differences. The human umbilical cord mesenchymal stem cells can be obtained rapidly and largely through the secondary culture to the primary culture system.
