Isolation and culture of bone marrow mesenchymal stem cells and their effects on immune regulation of CD8+T lymphocytes

doi:10.3969/j.issn.2095-4344.2017.05.005

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells have immunomodulatory properties and have potential applications in immunosuppression.
OBJECTIVE: To investigate the isolation and culture of bone marrow mesenchymal stem cells and the immunoregulation of CD8⁺T lymphocytes.
METHODS: Primary bone marrow mesenchymal stem cells were isolated from Wistar rats by bone marrow adherence method. The primary cells were purified by differential adherence and digestion, and collagen type I was added as extracellular matrix to expand bone marrow mesenchymal stem cells. Passage 2 bone marrow mesenchymal stem cells at densities of 0, 1×10³, 1×10⁴ and 1×10⁵/well were co-cultured with CD8⁺T lymphocytes, followed by phytohemagglutinin stimulation for 68 hours. T lymphocyte aggregation and proliferation were detected by staining with staining with 5,6-carboxyfluorescein diacetate succinimidyl ester and MTT, respectively.
RESULTS AND CONCLUSION: Different concentrations of bone marrow mesenchymal stem cells had an inhibitory effect on T lymphocytes, but this effect was weakest for the cells at the density of 1×10³ per well and strongest for the cells at the density of 1×10⁵ per well. Obvious agglomeration was observed in the same media. The proliferation rates of CD8⁺T lymphocytes were (44.83±4.92)%, (31.94±6.28)% and (15.77±3.98)%, respectively, after co-culture with 1×10³, 1×10⁴, 1×10⁵/well bone marrow mesenchymal stem cells. There were significant differences between groups (P < 0.05). These results showed that primary rat bone marrow mesenchymal stem cells could be obtained by direct adherence method of the whole bone marrow, and the cells could be purified by differential adherence combined with digestion and control method. Bone marrow mesenchymal stem cells could inhibit T lymphocyte proliferation in a concentration- dependent manner.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Mesenchymal Stem Cells, T-Lymphocytes, Tissue Engineering

CLC Number:

R394.2

Zhao Xiao-qiang, Shao Ming, Yang Hai-ping, Sun Ling. Isolation and culture of bone marrow mesenchymal stem cells and their effects on immune regulation of CD8⁺T lymphocytes[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(5): 682-686.

Figures/Tables 1

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