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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 observe and analyze the factors that affect the in vitro growth of mouse ColonyFormUnit Granulocyte and Monocyte (CFU-GM), and to establish a routine and stable CFU-GM culture method in our laboratory. Methods: Serum species, stimulating factor sources, agar preparation methods and BMC concentrations were counted and compared in culture system containing serum, stimulating factor, culture medium, agar and Bone MarrowCell (BMC) . Results: Under the same conditions, the colony yields of horse serum and newborn calf serum were 72 ± 12 9 and 34 ± 4 78 (P <0.01), respectively. The first cultured rat muscle conditioned medium and the second The colony yield of the second culture of rat muscle conditioned medium as stimulation factor group was respectively 58 52 ± 11 6 8 and 44 5 ± 7 46, P <0 01, and the supernatant was removed after cryopreservation and thawing The colony yields of the murine muscle conditioned medium and the murine muscle conditioned medium without the supernatant as stimuli were 42 ± 2 13 and 60 ± 12 6 49, respectively; P <0.01, formulated with triple distilled water Agar and D-Hank’s solution agar as support, the colony yields were 18 57 ± 333 and 42 5 ± 1 93, respectively, P <0.01; in the horse serum containing the first cultured rat muscle The correlation coefficient between the colony yield and the concentration of BMC implanted in the conditioned medium and the agar solution prepared by DHank’s solution was r = 0.91. CONCLUSION: Under the condition of constant BMC concentration and culture medium, horse serum was used as the serum and the first culture condition was stimulated. The agar was prepared with Danken’s liquid as the support, and the colony yield was the highest. ?