Transwell小室环境下人脂肪干细胞对不同年龄成纤维细胞功能的影响

Effect of human adipose-derived stem cells on function of human fibroblasts of different ages in Transwell system

目的:研究在Transwell小室环境条件下,脂肪干细胞(adipose-derived stem cells,ADSCs)分别对青年人来源和老年人来源成纤维细胞功能的影响及其差异。方法:从人脂肪组织中分离、培养ADSCs,从青年人和老年人皮肤组织中分离、培养青年人来源成纤维细胞和老年人来源成纤维细胞,用Transwell小室建立共培养模型,细胞增殖及细胞毒性检测试剂盒检测共培养后成纤维细胞增殖情况,RT-PCR检测共培养后成纤维细胞Ⅰ型胶原、基质金属激酶-1(matrix metalloproteinase-1,MMP-1)、β-半乳糖苷酶m RNA表达。结果:(1)共培养后老年人和青年人实验组成纤维细胞增殖分别高于老年人和青年人对照组成纤维细胞(P=0.000),且青年人实验组增殖高于老年人实验组(P=0.000)。(2)共培养后老年人和青年人实验组成纤维细胞的Ⅰ型胶原和β-半乳糖苷酶m RNA表达分别高于老年人和青年人对照组成纤维细胞(P=0.000),且青年人实验组的Ⅰ型胶原和β-半乳糖苷酶m RNA表达均高于老年人实验组(P=0.002、P=0.000)。(3)共培养后老年人和青年人实验组成纤维细胞的MMP-1 m RNA表达分别低于老年人和青年人对照组成纤维细胞(P=0.000),青年人实验组的MMP-1 m RNA表达低于青年人实验组(P=0.055),差异无统计学意义,此结果的产生可能与共培养后青年人成纤维细胞和老年人成纤维细胞增殖均较活跃,产生的MMP-1均较少有关。结论:ADSCs与成纤维细胞共培养,能促进不同年龄成纤维细胞的增殖能力和胶原蛋白的分泌能力。

Objective：To investigate the effect of adipose-derived stem cells（ADSCs） on young and elderly derived fibroblasts in Transwell system. Methods：ADSCs were isolated from human adipose tissues and were cultured. Fibroblasts derived from young people and old people were isolated from human skin tissue and were cultured. ADSCs and fibroblasts were co-cultured in Transwell chamber. Fibroblasts proliferation was evaluated by cell proliferation and cytotoxicity assay kit. Expression of fibroblasts collagen type I and matrix metalloproteinase-1 （MMP-1） and β-galactosidase mRNA were evaluated by RT-PCR. Results： （1）The proliferation of youth experimental group and elderly experimental group were obviously higher than that of young control group and elderly control group （P=-0.000）, and the proliferation of youth experimental group is higher than that of elderly experimental group （P=0.000）. （2）The expression of collagen type Ⅰ and β-galactosidase mRNA of youth experimental group and elderly experimental group was obviously higher than that of youth control group and elderly control group（P=-0.000）,and the expression of collagen type Ⅰ and β-galactosidase mRNA of youth experimental group was higher than that of elderly experimental group（P=-0.002,P=0.000）. （3）The expression of MMP- 1 mRNA of youth experimental group and elderly experimental group was obviously lower than that of youth control group and elderly control group（P=-0.000）, but the expression of MMP-1 mRNA of youth experimental group was lower than that of elderly experimental group（P=0.055）;the results may relate with the proliferation of fibroblasts and decreased production of MMP-1 in the elderly and the young after the co-culture. Conclusion：Fibroblasts cultured with ADSCs can promote fibroblasts proliferation and collagen secretion.