摘要
目的:探索骨髓间充质干细胞膜片的体外构建简便方法,并评估其成骨分化潜能。方法:将兔骨髓间充质干细胞高密度接种,在成骨诱导条件下连续培养形成细胞膜片。通过组织学、碱性磷酸酶活性定量检测、透射电镜和扫描电镜等方法,检测细胞膜片的特性。结果:高密度连续培养及成骨诱导后形成骨髓间充质干细胞膜片。HE染色证实膜片是一种由多层细胞和细胞外基质组成的聚集体;茜素红染色呈阳性;碱性磷酸酶定量分析含量较高;扫描电镜及透射电镜检查均见基质中大量的矿化结节聚集。结论:通过体外简单的连续培养和成骨诱导后,可形成具有良好成骨能力的骨髓间充质干细胞膜片。
Objective The convenient approach of constructing bone marrow mesenchymal stem cells （MSCs） sheets was explored. Methods We harvested MSCs sheets using a continuous culture method,Then,histological examination, quantitative analysis of ALP activity, scanning electron microscopy （SEM） and transmission electron microscopy （TEM） analysis were performed to evaluate the osteogenic sheets. Results After two weeks continuous culturing in vitro, MSCs sheets formed. HE staining confirmed sheets were composed of multiple layers of cells and their produced extracellular matrix. Furthermore, the cell sheets showed high ALP activities. The maintenance of the osteoblast specific phenotype was confirmed by positive staining for Alizarin red. In addition,both SEM and TEM analysis showed that distinct mineral- like nodules were observed. Conclusion Our study indicates a new technique of constructing MSCs sheets is established and the obtained sheets maintain the potential of osteogenic differentiation.
出处
《中国美容医学》
CAS
2012年第10期1776-1778,共3页
Chinese Journal of Aesthetic Medicine
基金
新疆维吾尔自治区自然科学基金项目(2012211A040)
关键词
间充质干细胞
细胞膜片
成骨分化
mesenchymal stem oells
cell sheet
differentiation
