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目的观察角化细胞生长因子(KGF)对体外小鼠骨髓间充质干细胞(MSCs)向肺泡上皮细胞诱导分化的影响。方法建立体外非接触共培养诱导小鼠MSCs向肺泡上皮细胞分化的实验模型,并在培养液中加入KGF(10μl/ml),小鼠分4组(6只/组),实验组A:MSCs(1×105)单独培养组+KGF;实验组B:MSCs+正常肺组织悬液(1×105)+KGF;实验组C:MSCs+损伤肺组织悬液+KGF;实验组D:MSCs+损伤肺组织悬液(1×105)。共同培养8 d,应用激光共聚焦显微镜和RT-PCR检测共培养体系下室MSCs中的SP-C和AQP5的表达情况。结果实验组C于共培养的d 8,较其它实验组的AQP5 mRNA的表达量明显增多(5550.00±244.39vs1650.17±184.02,1600.83±193.00,2890.00±179.09,P均<0.05);于共培养的d 5,与实验组A相比差异也有统计学意义(1743.17±228.41vs1482.00±156.11,P<0.05)。只有实验组C和D表达SP-C,且实验组C各培养时间点的SP-C的mRNA的表达量均较实验组D明显增多(2632.50±231.11vs1599.00±158.95;3976.67±221.81vs2066.83±101.31;5388.33±347.96vs3893.17±178.61,P均<0.01)。结论KGF可以进一步促进体外小鼠MSCs诱导分化为肺泡上皮细胞。
Objective To observe the effect of keratinocyte growth factor (KGF) on differentiation of mouse bone marrow mesenchymal stem cells (MSCs) into alveolar epithelial cells in vitro. Methods To establish an experimental model of MSCs differentiating into alveolar epithelial cells induced by non-contact co-culture in vitro. KGF (10μl / ml) was added into the culture medium. The mice were divided into 4 groups (6 mice / group) (1 × 105) cultured group + KGF alone; experimental group B: MSCs + normal lung tissue suspension (1 × 105) + KGF; experimental group C: MSCs + injured lung tissue suspension + KGF; experimental group D: MSCs + injured lung tissue Suspension (1 x 105). Co-cultured for 8 days, the expression of SP-C and AQP5 in the MSCs in the co-culture system was detected by laser confocal microscopy and RT-PCR. Results Compared with other experimental groups, the expression of AQP5 mRNA in experimental group C at d 8 was significantly increased (5550.00 ± 244.39 vs 1650.17 ± 184.02, 1600.83 ± 193.00, 2890.00 ± 179.09, P <0.05) D 5, which was also significantly different from that of experimental group A (1743.17 ± 228.41 vs 1482.00 ± 156.11, P <0.05). Only SP-C was expressed in experimental group C and D, and the expression of SP-C mRNA in experimental group C at each culture time point was significantly higher than that in experimental group D (2632.50 ± 231.11 vs1599.00 ± 158.95; 3976.67 ± 221.81 vs2066. 83 ± 101.31; 5388.33 ± 347.96vs3893.17 ± 178.61, P <0.01). Conclusion KGF can further promote the differentiation of mouse MSCs into alveolar epithelial cells in vitro.