Differences of human umbilical cord blood-derived mesenchymal stem cells cultured in different media

doi:10.3969/j.issn.2095-4344.2013.19.005

Abstract

Abstract:

BACKGROUND: The umbilical cord blood-derived mesenchymal stem cells are the hot spot in the field of stem cells, and there is no simple, effective culture method for the passage and amplification of umbilical cord blood-derived mesenchymal stem cells.
OBJECTIVE: To explore a better culture method of human umbilical cord blood-derived mesenchymal stem cells in vitro with different media in the separation of mesenchymal stem cells at a confluent status.
METHODS: Human umbilical cord blood was sterilely collected from full-term deliveries scheduled for cesarean section. They were assigned randomly into five groups: low-glucose culture medium group, high-glucose culture medium group, α-culture medium group, low-glucose culture medium + stem cell factor group, low-glucose culture medium + human marrow mesenchymal stem cells supernatant group. All culture medium used was Dulbecco’s modIfied Eagle’s medium. The cord blood mononuclear cells were isolated by lymphocyte separation medium. The monocytes of cord blood were inoculated into the culture medium containing 10% fetal bovine serum at 37 ℃ incubator with 0.05 volume fraction of CO2. Quantity and formation of cells were observed with invert microscope, and surface antigenic features were analyzed with flow cytometry.
RESULTS AND CONCLUSION: (1) Comparison on number of adherent cells and survival rate of mesenchymal stem cells cultured for 48 hours: Number of adherent cells in the low-glucose culture medium + human marrow mesenchymal stem cells supernatant group, and low-glucose culture medium + stem cell factor group were significantly increased (P < 0.05), while the survival rate of cells was also increased compared with low-glucose culture medium group, high-glucose culture medium group and α-culture medium group (P < 0.05). (2) Comparison on growth status of mesenchymal stem cells at different culture time points: Cell proliferation in the low-glucose culture medium + human marrow mesenchymal stem cells supernatant group, and low-glucose culture medium + stem cell factor group was more rapid than that in low-glucose culture medium group, high-glucose culture medium group and α-culture medium group at 3, 6, 9, 12, 15, 18 and 21 days (P < 0.05). There was no significant difference between low-glucose culture medium + human marrow mesenchymal stem cells supernatant group and low-glucose culture medium + stem cell factor group. Experimental findings indicate that, co-culture with low-glucose culture medium + stem cell factor or human marrow mesenchymal stem cells supernatant can promote the in vitro isolation, culture and proliferation of human umbilical cord blood-derived mesenchymal stem cells.

Key words: stem cells, umbilical cord and umbilical cord blood stem cells, umbilical cord blood-derived mesenchymal stem cells, ow-glucose Dulbecco’s modified Eagle’s medium, culture in vitro, stem cell factor, bone marrow mesenchymal stem cells, supernatant, isolation and culture, proliferation, stem cell photographs-containing paper

CLC Number:

R394.2

Zhao Xia, Lu Xi-jing, Liu Guo-qiang, Xu Min, Xing Jian, Wang Liang, Ding Hui-fang. Differences of human umbilical cord blood-derived mesenchymal stem cells cultured in different media[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(19): 3449-3454.

Figures/Tables 4

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