IL－6依赖细胞株的克隆化、冻存及其应用

Cloning,Cryogenic Storage and Application of IL-6 Dependent Cell Line

应用ＭＨ６０·ＢＳＦ２细胞检测ＩＬ－６生物学效价时，存在该细胞饲养困难和长期培养容易变异而失去ＩＬ－６增殖依赖性等问题。为此我们首先探讨了克隆化筛选ＭＨ６０·ＢＳＦ２的方法，从增殖依赖性较差的ＭＨ６０·ＢＳＦ２细胞群体中筛选到３株ＩＬ－６强依赖细胞株。并探讨了该细胞的冻存复苏方法。所冻存的细胞半年以上亦无ＩＬ－６增殖依赖性的改变。在此基础上采用ＭＴＴ比色法取代［￣３Ｈ］ＴｄＲ掺入法获得成功，从而使ＩＬ－６待测标本的生物活性检测更为方便经济。

In order to improve the method of IL-6 bicassay using IL-6 dependent cell line (MH60·BSF2).we es-tablished a cloning procedure and selected three clones of MH60·BSF2 whose proliferations were strongly dependent on IL-6,by comparison with uncloned MH60·BSF2. Meanwhile we stored the cloned MH60·BSF2 by cryogenic tech-nique. The cloned MH60·BSF2 cell line showed the same proliferating curve after cryogenic storage of half a year. We also developed the MTT coloy metric for MH60·BSF2 proliferation which showed the similar proliferating curve to the[￣3H]TdR method, The level of IL-6 of the culture supernant from different cells or cell line were measured. The results showed the PBMC of healthy individuals and of acute granulocyte leukemia secreted different value of IL-6(18u/ml and 234.67u/ml respectively);the human fetal gliocyets secreted high value IL-6(93.68u/ml).and LPS pro-moted the secretion of the gliocytes by about three folds.