RÉSUMÉ
Objective:To explore a method to obtain more mouse bone marrow cells more quickly.Methods:Take both lower limbs of the same mouse,the bone marrow cells were obtained by either the tube-nesting EP method or the traditional syringe irrigation method,recorded the time and quantity of bone marrow cells,to analyze the proportion of viable cells by flow cytometry,and to detect the proliferation of these bone marrow cells used CCK-8 method.Results:Compared with the traditional syringe irrigation method,the time of obtaining mouse bone marrow cells by tube-nesting EP method was shorter(P<0.000 1).Compared with the traditional syringe irrigation method,the number of bone marrow cells obtained by tube-nesting EP method was more(P<0.05).The proportion of live cells and proliferation ability of bone marrow cells obtained by the two methods were identical.Conclusion:Tube-nesting EP method is a new faster method to get more mouse bone marrow cells.
RÉSUMÉ
Objective To establish the cisplatin(DDP)-resistant cell line from human endometrial cancer cell line Ishikawa and to investigate its resistant mechanism to DDP. Methods A resistant endometrial cancer cell line ISH/DDP was established by gradually increasing dose of cisplatin and high-dose stimulation. The resistant index was estimated by 3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl)-2H-tetrazolium,inner salt (MTS) assay. Cell growth curve, doubling time and cell cycle phase distribution were measured; drug-resistant protein of breast cancer resistance protein (BCRP),P-glycoprotein (P-gp) and glutathione-S-transferase-π (GST-π) were examined by immuocytochemistry. Results The DDP-resistant cell line ISH/DDP was established with the resistant index of 3. 77. The proliferation of ISH/DDP got slow, doubling time prolonged, which were 40. 1 hours, while it was 34. 1 hours in Ishikawa (P0. 05] and G2/M phase [(11.9 ±0.7)% and (5. 7 ±2. 4)% ,P 0. 05) ,respectively. The score of the expression of GST-π in ISH/DDP and Ishikawa were 15. 2 ± 1. 9 and 14. 9 ± 1.1 (P > 0. 05) . Conclusion ISH/DDP cell line showed a typical resistant phenotype and biological characteristics, which may be accounted for high BCRP expression.