SRB法与MTT法细胞计数应用比较

Comparison between SRB way and MTT way applying to count cells in cells culture

目的比较SRB(SulforhodamineB)法与MTT法细胞计数的灵敏性与稳定性。方法取4种不同类型细胞系指数生长期细胞,调整细胞密度为1×106,并按比例稀释为0.078-10×105细胞梯度。分别用SRB法与MTT法测量各细胞梯度吸光度(A),分析其与相应细胞数目的相关性。SRB法与MTT法各随机取100个A值,测量其在2、16、24h和7d后的A值,比较这两种方法的稳定性。结果SRB法与MTT法测定的A值与不同细胞系细胞数目均有极好的相关性;SRB法的测定结果几乎不随时间而变,而MTT法的测定结果受时间影响较大。结论SRB法相对于MTT法是进行大规模细胞计数与细胞生长曲线测定前途更加光明的实验方法。

[Objective] To compare the susceptiveness and stability between Sulforhodamine B assay and MTT assay. [Method] The cells density of 4 cell line were adjusted to 1×10^6 by RPMI 1640. Then cells was diluted to a certain density in order as 5×10^5, 2.5×10^5, 1.25×10^5, 6.25×10^4, 3.13×10^4, 1.56×10^4 and 0.78×10^4. Then： The cells of every cells-density-gradient from every cell line were measured by Sulforhodamine B assay and MTT assay. The correlation coefficient between A value from Sulforhodamine B assay and MTT assay and the corresponding cell density was calculated, To compare the stability of A value beween two assay by assessing the A value variance beween A value from o hour and 2 hour, 16 hour, 24 hour or 7 days. [Result] As that from the MTT assay, the A value from the SRB assay was accord with the corresponding ceils density in all cell line .But compared with that from MTT assay, the A value from the SRB assay had less effects by the time variation. [Conclution] Compared with the MTT assay, the SRB assay is a better way to calculate growth curve and to estimate the ceils density when the sample size is very large.