人胰岛素样生长因子-1基因强化组织工程法诱导裸鼠异位软骨形成

Ectopic Chondrogenesis of Nude Mouse Induced by hIGF-1 Gene Enhanced Tissue Engineering Technology

目的探讨人胰岛素样生长因子-1(hIGF-1)基因转染诱导后的人骨髓基质干细胞(hMSCs)与再生丝素膜复合后在裸鼠体内形成组织工程软骨的能力。方法分离培养hMSCs,体外诱导成软骨样细胞,流式细胞仪测定细胞表型,免疫组化检测软骨细胞表型;脂质体介导hIGF-1转染诱导后的hMSCs,RT-PCR、酶免疫测定法和免疫组化检测hIGF-1及Ⅱ型胶原的表达;转染后的hMSCs与再生丝素膜复合培养,扫描电镜观察细胞形态,复合物植入裸鼠皮下,HE及免疫组化染色观察新生软骨的结构和表型。结果分离培养出纯化的hMSCs符合间充质干细胞表型,具有快速增殖及经诱导后向软骨分化的能力,诱导后的细胞表达Ⅱ型胶原;转染诱导后的hMSCs能稳定分泌具活性的hIGF-1蛋白,并表达软骨细胞表型;转染后的hMSCs与再生丝素膜复合培养显示良好的生物相容性,新生组织具典型的软骨组织陷窝,并表达Ⅱ型胶原。结论经hIGF-1基因诱导后的hMSCs与再生丝素膜在体内能构建出软骨样组织。

Objective To duced by human insulin like explore growth the capacity of ectopic chondrogenesis of nude mouse in-duced by human insulin like growth favtor-1 （hIGF-1）gene enhanced tissue engineering technology. Methods Human mesenchymal stem cells （hMSCs） were isolated, cultivated and induced to differentiate towards chondrocytes with its phenotypes assayed by flow cytometer and immunohisto- chemical method; After transferred by means of Lipofectamine, RT-PCR,EIA and immunohistoche- mistry to determine the expression of＇ the induced hMSCs hIGF-1 and type-Ⅱ collagen; The complexes of hMSCs-hIGF-l-regenerated silk fibroin film were observed under scanning electron micro-scope and examined histologically and immunohistchemically after being implanted under the dorsal skin of nude raise. Results HMSCs showed potentials for proliferation and chondrogenesis differentiation with similar characteristic of MSCs and cartilages. The induced hMSCs after being transferred showed expression of hIGF-1 and type-Ⅱ collagen. The transferred hMSCs show well compatibility on the surface of regenerated silk fibroin film. The implanted complexes showed formation of new cartilage. Conclusion Ectopic chondrogenesis can be induced by hIGF-1 gene enhanced tissue engineering technology.