摘要
目的 :观察分析影响小鼠粒单系祖细胞 (ColonyFormUnit GranulocyteandMonocyte ,CFU GM )在体外生长的因素 ,建立本实验室常规 ,稳定的CFU—GM培养方法。方法 :在含血清、刺激因子、培养液、琼脂和骨髓有核细胞 (BoneMarrowCell,BMC)的培养体系中 ,分别变换血清种类、刺激因子来源、琼脂配制方法和BMC浓度 ,计数、比较祖细胞集落。结果 :在其它条件相同时 ,马血清和新生小牛血清组的集落产率分别为 72± 12 9和 34± 4 78,P <0 0 1;第一次培养的鼠肌条件培养液和第二次培养的鼠肌条件培养液作为刺激因子组的集落产率分别为 5 8 2 5± 11 6 8和 44 5± 7 46 ,P <0 0 1,经冻存并自然融化后去除上清液的鼠肌条件液和未去除上清液的鼠肌条件液作为刺激因子组的集落产率分别为 42± 2 13和 6 0± 12 6 49;P <0 0 1,以三蒸水配制的琼脂和D -Hank’s液配制的琼脂作为支撑物 ,集落产率分别为 18 5 7±3 33和 42 5± 1 93 ,P <0 0 1;在含马血清、第一次培养的鼠肌条件液和D Hank’s液配制的琼脂的体系中 ,集落产率与所种入BMC的浓度的相关系数r =0 .91。结论 :在BMC浓度和培养液不变的条件下 ,以马血清为血清 ,第一次培养的肌条件液为刺激因子 ,D Hank’s液配制的琼脂为支撑物 ,集落产率最高 ,且方法稳?
Objective:To obsevate and analyse the impact factors witch effect on the growth of CFUGM in culture on vitro, further make the stable method of CFUGM culture in our labratory.Methods:The serum.Colony stimulating factor、culture medium、agar and bone marrow cells made up of culture system, changed the sort of serum、the making way of CSF、the way of agar dilated and the concetration of BMC in system,then culture for 7 days and count the numbers of CFU-GM.Results:The numbers of CFUGM are 72±12.9 in the group of horse serum and 34±4.78 in the group of newborncalf serum, P<0.01; in the group of mouse muscle conditioned medium witch get from the mouse muscle cultured only once as CSF the numbers of CFU-GM are 58.25±11.68, and are 44.5±7.46 in the group of mouse muscle cultured twice, the P<0.01; in the group of mouse muscle conditioned medium which freezed and melted in the room temprature, the numbers are 60±12.64, when throw out the supernatant of melted liquid, the numbers are 42±2.13, the P<0.01; in the group of agar diluted with trippl distilled water and with Dhanks buffer are 18.57±3.33 and 42.5±1.93 respectively, P<0.01. In the culture system witch include horse serum, the mouse muscle conditioned medium cultured only once. the agar diluted by DHanks buffer liquid, the correlation coefficient betweet the numbers of CFUGM and the concetration of BMC is 0.91.Coclusion:the numbers of CFUGM are most and stable. in the culture system contains horse serum. mouse muscle conditioned medium cultured once and agar diluted by DHanks buffer.
出处
《四川省卫生管理干部学院学报》
2000年第2期81-83,共3页
Journal of Sichuan Continuing Education College of Medical Sciences
基金
学院科研基金课题
