Method validation of lymphocyte subgroup phenotyping analysis in cynomolgus monkeys and Sprague-Dawley rats using flow cytometry
Method validation of lymphocyte subgroup phenotyping analysis in cynomolgus monkeys and Sprague-Dawley rats using flow cytometry作者机构：Center for Drug Safety Evaluation and ResearchShanghai Institute of Materia MedicaChinese Academy of Sciences
出 版 物：《中国药理学与毒理学杂志》 (Chinese Journal of Pharmacology and Toxicology)
年 卷 期：2013年第3期
页 面：513-513页
核心收录：
学科分类：1007[医学-药学(可授医学、理学学位)] 1006[医学-中西医结合] 100706[医学-药理学] 100602[医学-中西医结合临床] 10[医学]
主 题：lymphocyte phenotype flow cytometry
摘 要：OBJECTIVE To validate a flow-cytometry-based method for the measurement of lymphocyte subgroups in cynomolgus monkeys and Sprague Dawley rats,*** For cynomolgus monkeys,two panels of fluorescent antibodies,namely CD3-FITC /CD4-PerCP /CD8-PE and CD3-PE-Cy5.5 /CD16-PE /CD20-FITC were used to analyze the proprotion of T cells including T helper cell(CD3 + CD4 +) and T cytotoxic cells(CD3 + CD8 +),NK cells(CD3 CD16 +) and B cells(CD3-CD20 +).For Sprague Dawley rats,other two panels of fluorescent antibodies,namely CD3 FITC /CD4-PE /CD8a-PerCP and CD3-FITC /CD45RA-PE-Cy5.5 /CD161a-PE were used to analyze the proprotion of T cells,NK cells(CD3-CD161 +) and B cells(CD3-CD45RA +).Blood samples from 10 monkeys and 10 rats were analyzed for intra-assay precision,sample stability and inter-subject ***,intra-animal precision was investigated in *** For cynomolgus monkeys,intra-assay CV and intra-animal CV for all parameters were within 15%.The positive binding signals of all isotype controls were less than 1%.Furthermore,monkey blood samples were stable for all parameters when kept at room temperature for 6 h or at 4℃ for 1 *** after fixation were stable while stored in the refrigerator(+ 4℃) for 24 *** rats,intra-assay CVs for all parameters were within 15%.The positive binding signals of isotype controls of CD3 + /CD4 + and CD3 + /CD8 + were less than 1%.Furthermore,rat blood samples were stable for CD3 + /CD4 + and CD3 + /CD8 + while stored in the refrigerator(+ 4℃) for 24 ***,rat blood samples were unstable for CD3 /161a when kept at room temperature for 6h and unstable for CD3 /45RA when kept at 4℃ for 1 *** latter two cases,the positive binding signals of the isotype controls were around 2%.CONCLUSION All the parameters tested fulfilled the criteria of acceptance with a few minor exceptions related to stability data from rat blood *** is suitable for the measurement of lymphocyte subgroups in cynomolgus mo
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