MRC-5人二倍体细胞培养条件的优化

The optimization of culture conditions of MRC-5 human diploid cell

目的对MRC-5人二倍体细胞培养条件的优化比较。方法用3种不同的培养基将MRC-5人二倍体细胞在T25方瓶和Spinner培养系统Cytodex1微载体2种培养体系进行培养比较,每天观察细胞形态,进行细胞计数,绘制生长曲线,并检测葡萄糖-乳酸值,筛选出1种较适宜培养基,用不同级别的牛血清比较其生长增殖情况。结果 3种培养基在细胞形态,细胞的贴壁、分裂以及维持等方面无明显差异。但在增殖上,在2种培养体系中MEM(43.25±0.60)×104cells/m L,(12.98±1.27)×105cells/m L与M199(35.40±1.41)×104cells/m L,(10.76±1.31)×105cells/m L及DMEM/F12(36.75±1.59)×104cells/m L,(11.22±1.42)×105cells/m L比较,差异均有统计学意义(P<0.01),MEM培养液细胞增殖5.17和6.49倍,优于M199和DMEM/F12培养液。进口胎牛血清细胞增殖(4.55±0.51)×105cells/m L,优于其他3种牛血清的(4.12±1.03)×105cells/m L、(3.59±0.48)×105cells/m L和(3.53±0.52)×105cells/m L。结论 3种培养基都可以用于MRC-5人二倍体细胞的繁殖培养,但MEM培养液更佳。进口胎牛血清较其他血清有优势。

Objective To optimize the culture conditions of MRC- 5 human diploid cell. Methods To compare the growth status of MRC- 5 cells,three kinds of culture medium with T25 bottles and Spinner cultivation system Cytodex1 micro carrier were used. Morphology,cell counting,growth curve,glucose- lactic acid value were observed and detected daily for screening a kind of suitable medium. Cell proliferation was compared with different levels of the bovine serum. Results There were no significant differences among the three kinds of culture medium. There were significant differences among MEM（（ 43. 25 ± 0. 60） × 10^4 cells / m L,（ 12. 98 ± 1. 27） × 10^5 cells / m L）,M199（（ 35. 40 ± 1. 41） × 10^4 cells / m L,（ 10. 76 ± 1. 31） × 10^5 cells / m L） and DMEM / F12（（ 36. 75 ± 1. 59） × 10^4 cells / m L,（ 11. 22 ± 1. 42） × 10^5 cells / m L）（ P 0. 01）. The cell proliferation of MEM cultures was 5. 17 and 6. 49 times better than those of M199 and DMEM / F12 cultures. Imported fetal bovine serum cell proliferation（（ 4. 55 ± 0. 51） × 10^5 cells / m L） was better than the other three bovine serum（（ 4. 12 ± 1. 03,3. 59 ± 0. 48,3. 53 ± 0. 52） × 10^5 cells / m L）. Conclusion Tree kinds of culture medium can be used to culture MRC- 5 human diploid cell. The MEM culture is better. Imported fetal bovine serum is better than other kinds of serum.