酸性磷酸酶法检测体外培养细胞数

利用小鼠成纤维细胞系(NIH3T3)、小鼠骨髓瘤细胞系(SP2/0)、人大肠癌细胞系(LO-VO)和人白血病细胞系(K562),评价酸性磷酸酶(APA)法用于检测体外各类型细胞的增殖和杀伤作用。用直线回归分析光吸收度与每孔活细胞数的关系。结果表明,APA法能准确地反映检测的活细胞数(相关系数均>0.99)。本方法不仅能很好地检测表皮生长因子对细胞的增殖作用,也能够检测顺铂对体外细胞的杀伤作用。结果表明APA法简单、灵敏,可以用于上皮和间质等贴壁和悬浮生长的细胞计数。

ACID PHOSPHATASE ASSAY (APA) FOR DETERMINATION OF CELL NUMBER IN CULTURE

An acid phosphatase assay (APA)for determination of the numbers of various cell types in culture was evaluat-ed. 4 cell lines were used,including NIH3T3,SP2/0,LOVO(human colon cancer cell line)and K562(human leukemic cell line). The celis were seeded into 96-well culture plates in different density ,which were measured by APA,and lin-er aggression was used to test the relationship of absobrance and cell number per well. The results showed that the relationship between cell number per well and absorance determined by APA was excellent (T>0. 99 in 4 cell lines). Al-so,the proliferative curve of LOVO celis stimulated by epidermal growth factor and killing curve of LOVO cells treat-ed with cisplatin were precisely recorded using APA. It was found that the assay could detect at least 25. 80 cells per well , and reaction times recommended for APA should be between 2 - 3 hours. It is suggested that APA is a rapid, simple,precise and sensitive method for determination of cell number,including adherent and non adherent cells.