巨噬细胞条件培养液对原代培养小鼠成肌细胞生长的影响

目的 观察小鼠巨噬细胞条件培养液对小鼠成肌细胞生长的影响.方法 小鼠腹腔内连续3d注射肌匀浆和淀粉混合物,然后分离小鼠腹腔巨噬细胞,在无血清培养液中培养48h后收集上清.取生后5d小鼠的成肌细胞,在含10%血清的DMEM/F12培养液中培养36～48h后,将血清浓度降低至0.5%.将巨噬细胞培养上清加入低浓度血清培养的成肌细胞中,作用72h后收集细胞,通过Wright染色、LDH细胞化学和免疫细胞化学方法观察成肌细胞的生长情况.结果 巨噬细胞条件培养液能够在低浓度血清的条件下维持成肌细胞的正常形态和活性,并促进成肌细胞增殖.实验组LDH阳性细胞与对照组相比,其平均吸光度与积分吸光度均存在极显著性差异(分别为P＜0.001和P＜0.000 1).免疫细胞化学显示,实验组desmin阳性细胞数明显多于对照组.结论 巨噬细胞条件培养液中含有促进小鼠成肌细胞生长的因子.

THE EFFECT OF MACROPHAGE-CONDITIONED MEDIUM ON THE GROWTH OF PRIMARY MICE MYOBLASTS

Objective To investigate the effect of macrophage-conditioned medium on the growth of mouse myoblasts. Methods After a 3-day intraperitoneal injection with a mixture of muscle homogenate and starch,macrophages were obtained from mouse peritoneal cavity and cultured for 48 hours in serum-free DMEM/F12.Then the cultured medium was harvested.Myoblasts were obtained from newborn mice skeletal muscle and grown in DMEM/F12 supplied with 10% FBS for 36-48 hours,and then exposed to 0.5% FBS medium with or without macrophage-conditioned medium.Cells were harvested at 72 hours later,their growth was observed with Wright staining,LDH cytochemistry and immunocytochemistry. Results Macrophage-conditioned medium can maintain the normal morphology and the activity of myoblasts grown in low concentration of serum and promote their proliferation.There was a significant difference between average absorbance and integral absorbance of LDH positive cells of the two groups(P<0.01 and P<0.000 1 respectively).The experiment group contained more positive cells compared with the control group,identified by immunocytochemistry for desmin.Conclusion Macrophage-conditioned medium contains factors that promote growth of mice myoblasts.