摘要
目的 细胞与细胞之间及细胞与基质之间的粘附作用是细胞间信号传递及多种细胞生物活动的基础 ,细胞粘附作用的检测是细胞生物学研究中重要的实验方法之一。本文对采用51Cr释放实验及3 H -TdR掺入法检测细胞粘附的方法进行了比较。方法 将铺底细胞以 0 5× 10 5 /孔的密度于 96孔培养板中铺底培养 2 4h ,待检细胞以51Cr或3 H -TdR标记 ,以 1× 10 5/孔的浓度加入96孔培养板中 ,作用 4h后轻柔洗去未粘附细胞。按相应检测系统检测51Cr及3 H -TdR标记的粘附细胞的放射强度 ,比较其灵敏度和稳定性。结果 51Cr释放实验及3 H -TdR掺入法均具有较好的敏感性和稳定性。相比较 ,3 H -TdR掺入法的敏感性和稳定性更好。这两种方法均能准确地检测细胞间的粘附现象 ,并反映其相对强弱。结论 采用同位素标记的方法 ,其敏感性和稳定性均较好 ,且干扰因素少 ,所获结果稳定可靠 ,3 H -TdR掺入法的敏感性和稳定性较51Cr释放法好。
Objective To compare the two cell adhesion assay techniques based on 51 Cr release and 3H-TdR incorporation.Methods Firstly,the cells to be tested were cultured to confluence in the 96 well plate for 24 hours. After with 51 Cr or 3H-TdR label, the isotope labeled cells were add into plate wells and incubated for another 4 hours. Then the un-adhered cells were removed by gently washing. The cpm of two assay system were counted, the sensitivity and stability of two methods were compared.Results Assay methods based on 51 Cr release and 3H-TdR incorporation could both reflect the cell adhesion level correctly. In assaying sensitivity and stability showed that the 3H-TdR incorporation assay was better than in 51 Cr release assay.Conclusions Adhesion method based on isotope label could provide good sensitivity and stability. The sensitivity and stability of 3H-TdR incorporation is better than that of 51 Cr release assay.
出处
《中国医师杂志》
CAS
2003年第1期7-9,共3页
Journal of Chinese Physician
基金
国家自然科学基金资助项目 (3970 0 65)
