三种结缔组织体外诱导胎儿骨髓间充质干细胞的分化

Differentiation of fetal-bone mesenchymal stem cells induced by three kinds of connective tissue in vitro

目的比较胎儿肠黏膜下层、肠系膜与羊膜的结缔组织体外诱导胎儿骨髓间充质干细胞(BMSCs)分化为上皮细胞作用的差异及其可能机制。方法制备去上皮的3种结缔组织块,检测其中有无EGF和IGF-1表达。将4,6-联脒-2联苯基吲哚(DAPI)标记的P3-BMSCs滴加于3种结缔组织上培养,设结缔组织诱导组、结缔组织与表皮生长因子(EGF)和胰岛素样生长因子-1(IGF-1)联合组、EGF、IGF-1诱导对照组。于培养第8、10和12d收集组织块,比较各组诱导BMSCs向上皮细胞分化的差异。结果经3种结缔组织和生长因子诱导后的BMSCs均表达CK和EMA;联合诱导组作用强于单用结缔组织组(P<0.05);3种结缔组织中以羊膜的作用最强(P<0.05)。3种组织均表达EGF,肠黏膜下层及肠系膜还表达IGF-1。结论胎儿肠黏膜下层、肠系膜与羊膜的结缔组织以及EGF、IGF-1在体外均有诱导BMSCs分化为上皮的作用;EGF和IGF-1可协同增强3种结缔组织的诱导作用;结缔组织中的EGF或IGF-1可能是其诱导作用的内源性因素。

Objective To compare the inducing effect of three kinds of connective tissue from anmion, fetal mesentery and intestinal submucosa on the differentiation of fetal BMSCs in vitro. Methods The epithelium on three kinds of connective tissue were sampled and EGF and IGF-1 in these tissue clumpes were detected. DAPI-P3-BMSCs were planted on the surface of three kinds of connective tissue to culture. There were connective tissue induced groups, connective tissue and EGF and IGF-1 induced groups, EGF and IGF-1 induced as control group. The tissue clump was collected at 8 d,10 d,12 d, to compare their inducing effect on the differentiation of BMSCs. Results All of BMSCs induced by three kinds of connective tissue and growth factors expressed CK or EMA. The effect of unite-induced groups was stronger than those groups induced by only connective tissue, P 〈 0. 05 ; The strongest inductive effect of three connective tissue was exerted by anmion, P 〈 0. 05. EGF was expressed in the three kinds of connective tissue, and IGF-1 was only expressed in mesentery and intestinal submucosa. Conclusion BMSCs could be induced to differentiate into epithelial cells by the connective tissue of amnion, mesentery and intestinal submucosa, EGF and IGF-1 in vitro; EGF, IGF-1 could increase the effects of these connective tissue on the induction of BMSCs differentiation; EGF and IGF-1 may be one of the endogenic factor in these connective tissue which induces BMSCs to differentiate into epithelium.