羊膜促进间充质干细胞表皮细胞生长的形态学特征

The investigation of pathology about the reconstruction of skin with bone marrow-derived mesenchymal stem cells and keratinocyte

目的将人羊膜(humanamnioticmembrane,HAM)负载培养猪骨髓间充质干细胞(MSCs)重建皮肤真皮替代物,负载培养猪表皮细胞重建皮肤表皮替代物,并观察猪MSCs、表皮细胞在HAM上生长增殖的形态特点。方法将贵州小香猪的MSCs、表皮细胞原代培养,传代扩增后,分别以不同的密度接种于HAM的基质面,逐日于倒置显微镜下观察MSCs、表皮细胞的生长增殖变化情况,并于培养3～18d进行光镜、电镜观察,检测MSCs、表皮细胞在HAM上生长的形态特点。结果接种后30min内就明显见到MSCs、表皮细胞在HAM上贴附,24h内大部分贴附生长,3d形成单层完全覆盖HAM,超微结构形态良好。结论HAM是MSCs、表皮细胞在体外培养的良好载体,对MSCs、表皮细胞有明显的促增殖作用。

Objective Reconstructing dermis and epidermis equivalent by means of human amniotic membrane(HAM)loading respectively bone marrow derived mesenchymal stem cells (MSCs) and keratinocytes of piglet with tissue engineering technology, and observing the morphological characteristics of HAM loading MSCs and keratinocytes respectively. Methods The primary cultures of MSCs and keratinocytes of Guizhou piglet, subsequently MSCs and keratinocytes were expanded and bred by means of different densities on the stroma surface of HAM for many times. The growth and proliferation of MSCs and keratinocytes were observed under invert microscope each day. From 3 days to 18 days after cultured, the changes of growths of MSCs and keratinocytes on HAM were examined under light microscope and electron microscope. Results It was evident that MSCs and keratinocytes adhere to the stroma surface of HAM after cultured about 30 minutes. Most of MSCs and keratinocytes were seen adhering to the stroma surface of HAM within 24 hours.MSCs and Keratinocytes formed monolayer and covered completely HAM within 3 days. The ultrastructure of HAM loading MSCs and keratinocytes was well. Conclusion HAM is a good carrier of MSCs and keratinocytes cultured on it in vitro. HAM is able to promote the proliferation of MSCs and keratinocytes significantly.