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在测定大鼠肉瘤(XC)细胞和莫里斯肝癌7777(MH)细胞的非锚定依赖性细胞生长中,3H-胸腺嘧啶核苷掺入法与集落形成试验的比较

3H-thymidine incorporation method versus colony forming assay in the determination of anchorage-independent cell growth of rat sarcoma (XC) and Morris hepatoma 7777 (MH) cells.

作者信息

Klein A, Madeja Z, Guzik K

机构信息

Department of Animal Biochemistry, Jagellonian University, Kraków, Poland.

出版信息

Folia Histochem Cytobiol. 1991;29(2):75-80.

PMID:1804725
Abstract

The modification of 3H-thymidine incorporation method of Tanigawa was used to the estimation of anchorage-independent growth of virally and chemically transformed rat cells. The relationship between colony forming assay and 3H-TdR incorporation test was determined, depends on the composition of culture medium and the period of incubation of rat sarcoma (XC) cells with thymidine. The influence of exogenous mitogens (RFG, TGF beta 1 and insulin) and autocrine factor (at different step of purification) on the growth of Morris hepatoma 7777 (MH) cells was estimated by both methods. Regression analysis comparing the results of colony counting and thymidine incorporation revealed good correlation between the two methods. The modification can be used the determination of growth stimulating or growth inhibiting activity and in multistep purification procedure of autocrine growth factors.

摘要

采用谷川的³H-胸腺嘧啶核苷掺入法的改进方法来评估病毒转化和化学转化的大鼠细胞的非贴壁依赖性生长。确定了集落形成试验与³H-TdR掺入试验之间的关系,这取决于培养基的组成以及大鼠肉瘤(XC)细胞与胸腺嘧啶核苷孵育的时间。通过这两种方法评估了外源性有丝分裂原(RFG、TGFβ1和胰岛素)和自分泌因子(在不同纯化步骤)对莫里斯肝癌7777(MH)细胞生长的影响。比较集落计数和胸腺嘧啶核苷掺入结果的回归分析表明这两种方法之间具有良好的相关性。该改进方法可用于确定生长刺激或生长抑制活性以及自分泌生长因子的多步纯化过程。

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