小鼠白细胞介素-3的制备及其检测条件的分析

STUDIES ON OPTIMAL CONDITIONS FOR MOUSE IL-3 PRODUCTION AND DETECTION

从小鼠白细胞介素-3(IL-3)分泌细胞株WEHI-3获取了IL-3,同时用小鼠IL-3依赖细胞株32-D检测IL-3和滴定标本中IL-3的活性单位,并对检测条件进行了探索。结果表明,WEHI上清中含高活性IL-3,不含IL-2,其最佳收集条件是5×10~5/ml细胞浓度培养48h。32-D细胞生长依赖于IL-3。检测细胞的浓度、培养时间及同位素加入时间直接影响检测结果,最佳检测条件是(5～10)×10~5/ml细胞浓度、培养36h、终止前12h加~3H-TdR。

Conditioned Media from mouse IL-3 producing cell line WEHI-3 (WEHI-OM) were collected and IL-3 activity measured in a proliferation response system using the IL-3 dependent cell line 32-D, Optimal conditions for IL--3 production and detection were studied. Results indicated that WEHI-OM contained high levels of IL-3, but no IL-2. Supernatants showed highest IL-3 activity when cultured with WEHI-3 cells at 5×105/ml concentration for 48 hrS. The results also showed that 32-D cells were completely dependent on IL-3. The best condition for detection of IL-3 activity was for 32-D cells to be incubated at(5-10)×105 /ml concentration for 24 hrs with subsequent addition of 3H-thymidine 12 hrs before harvesting.