Isolation and culture of bone marrow mesenchymal stem cells and their effects on immune regulation of CD8+T lymphocytes / 中国组织工程研究

ABSTRACT

BACKGROUND:Bone marrow mesenchymal stem cel s have immunomodulatory properties and have potential applications in immunosuppression. OBJECTIVE:To investigate the isolation and culture of bone marrow mesenchymal stem cel s and the immunoregulation of CD8+T lymphocytes. METHODS:Primary bone marrow mesenchymal stem cel s were isolated from Wistar rats by bone marrow adherence method. The primary cel s were purified by differential adherence and digestion, and col agen type I was added as extracel ular matrix to expand bone marrow mesenchymal stem cel s. Passage 2 bone marrow mesenchymal stem cel s at densities of 0, 1×103, 1×104 and 1×105/wel were co-cultured with CD8+T lymphocytes, fol owed 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 cel s had an inhibitory effect on T lymphocytes, but this effect was weakest for the cel s at the density of 1×103 per wel and strongest for the cel s at the density of 1×105 per wel . 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/wel bone marrow mesenchymal stem cel s. There were significant differences between groups (P<0.05). These results showed that primary rat bone marrow mesenchymal stem cel s could be obtained by direct adherence method of the whole bone marrow, and the cel s could be purified by differential adherence combined with digestion and control method. Bone marrow mesenchymal stem cel s could inhibit T lymphocyte proliferation in a concentration-dependent manner.