新型淋巴细胞培养基配制及其性能比较评价

Preparation and performance evaluation of a new culture medium for lymphocyte

目的比较大鼠淋巴细胞在不同培养基中的生长及增值状况,筛选最佳新型淋巴细胞培养基。方法分离培养大鼠胸腺淋巴细胞,分别采用新配方培养基、无血清RPMI-1640培养基和无血清新配方培养基培养细胞,用倒置显微镜观察细胞的生长状态,MTT比色法检测细胞增殖活性。结果镜下观察,接种24 h后不同培养基中细胞均匀大量贴壁并伸展,5 d细胞生长数量减少,出现衰退;含0.1 mmol的蛋氨酸脑非肽(MEK)的新型淋巴细胞基可大幅度提高淋巴细胞生长,细胞生长状态良好,新型培养基中淋巴细胞增殖活性与传统RPMI-1640比较差异均有统计学意义(P<0.05);在RPMI1640培养基、新配方2培养和无血清新配方2培养基中淋巴细胞增殖活性分别为99.9%、103.1%和96.6%。结论新型培养基较RPMI-1640更适合淋巴细胞生长及增殖。

Objective To compare the growth and proliferative status of rat lymphocytes in different media, and to screen the new medium for lymphocyte culture. Methods Isolated and cultured rat thymus lymphocytes were cultured in a new medium, serum-free RPMI-1640 medium and a new serum-free medium. The growth states of the cells were observed with inverted microscope. The cell proliferative activity was detected by 3- （ 4,5-dimethylthiazol-2-yl ） -2,5-di- phenyltetrazolium bromide（MTT）colorimetric assay. Results Twenty-four hours after the inoculation, a large number of cells in different media adhered to the wall and spread evenly. Five days after the inoculation the number of cell growth was reduced, and the cells were declined. The new medium containing 0. 1 mmol of methionine-enkephalin （MEK） could improve lymphocyte cell growth greatly. There was a significant difference in the cell proliferative activity between the new medium and traditional RPMI-1640 medium （P 〈 0. 05 ）. The lymphocyte proliferation activity in RPMI-1640, the new medium（ formulation 2 ） with serum, and the serum-free new medium （ formulation 2 ） was 99. 9% , 103.1%, and 96. 6% ,respectively. Conclusion The new medium prepared is more suitable for the growth and proliferation of rat lymphocyte.