关键词： 溴化四唑蓝，重组人粒细胞集落刺激因子，重组人粒细胞／巨噬细胞集落刺激因子，重组人肿瘤坏死因子，生物学活性，(k,k)法

摘要：目的：对MTT法测定重组人粒细胞集落刺激因子(rhG-CSF)、重组人粒细胞／巨噬细胞集落刺激因子(rhGM-CSF)、重组人肿瘤坏死因子(rhTNF-α)等基因工程细胞生长因子生物学活性进行方法学研究。方法：分别培养NFS-60、L929、TF-1细胞，用MTT进行染色后，用450型酶标读数仪在570mm处测定OD值。结果：确定了不同细胞株对MTT的最佳染色浓度、染色时间和回归关系及线性范围，找出了用MTT法测定rhG-CSF、rhGM-CSF、rhTNF-α等基因工程细胞生长因子生物学活性的灵敏度范围及线性回归关系。结论：为将(k,k)法用于MTT法测定基因工程细胞生长因子生物学活性奠定了基础。

The Determination for Biological Activities of Three Recombined Human Cell Growth Factors by MTT Method

jia Qian,Zhou Xingjun,Wang Hui,Wang Yanfang,Tan Lixia,Zhang Weiting(R D Center of North China Pharmaceutical Corporation(Shijiazhuang 050015))

Abstract：It was investigated that the biological activities of rhG-CSF,rhGM-CSF and rhTNF-α were determined by MTT method.Methods:Culture the NFS-60、L929 and TF-1cell lines respectively and measure the OD value by EIA reader on 570nm after dyed with MTT.Results:The optimized dying concentration,dying time,regression relation and linear region for different cell lines were found.Furthermore the sensitivity and the linear regression of the biological activity of rhG-CSF、rhGM-CSF and rhTNF-α determined by MTT method were obtained.Conclusion:All these will constitute the basis for(k,k)method application in the determination of biological activities of cell growth factors by MTT.

Keywords：MTT,rhG-CSF，rhGM-CSF，rhTNF-α,Biological activity,(k,k)method参考文献：

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收稿日期：1999-07-13

修稿日期： 1999-09-24