MTT法测定三种基因工程细胞生长因子生物学活性方法研究

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

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 6 0、L92 9、TF 1细胞 ,用MTT进行染色后 ,用 45 0型酶标读数仪在 5 70mm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)

Purpose:It was investigated that the biological activi t ies of rhG-CSF,rhGM-CSF and rhTNF-α were determined by MTT method.Meth ods: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:T he optimized dying concentration,dying time,regression relation and linear regio n 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 dete rmined by MTT method were obtained.Conclusion:All these will constitut e the basis for (k,k) method application in the determination of biological acti vities of cell growth factors by MTT.