人牙周膜干细胞膜片的制备

Preparation of human periodontal ligament stem cell sheet

目的制备人牙周膜干细胞(h PDLSC)膜片。方法刮取接受正畸治疗的20例患者完整拔除的前磨牙牙周膜组织,机械分离加酶消化法分离培养人正常牙周膜细胞,采用有限稀释法克隆培养纯化h PDLSC,测算克隆形成率,检测h PDLSC的体外多向分化潜能。向培养基中添加40μg/m L维生素C制备h PDLSC膜片,用扫描电镜及HE染色观察所制备的h PDLSC膜片的结构。结果 h PDLSC具有较高的克隆形成率,体外诱导具有成骨成脂多向分化潜能。HE染色显示h PDLSC膜片由3、4层细胞构成,细胞与细胞之间连接紧密,富含细胞外基质。扫描电镜显示h PDLSC膜片中h PDLSC排列紧密,细胞外基质丰富。结论成功制备了h PDLSC膜片。

Objective To prepare the human periodontal ligament stem cell（ h PDLSC） sheet. Methods Human periodontal ligament stem cells were isolated and cultured from 20 extracted premolar teeth. ALP staining and red oil O staining were utilized to observe the differential ability of h PDLSC. h PDLSC sheet was prepared by adding 40 μg/m L vitamin C into the culture medium,HE and SEM were used to observe the cell sheet structure. Results In this study,h PDLSC was successfully isolated and cultured; it had high colonic ability and multiple differentiation ability which complied with the characteristics of mesenchymal stem cells. SEM indicated that the cells were arranged closely,and the extracellular matrix was abundant. HE staining showed that the cell sheet was composed of 3 to 4 layers of cells. Cells and cells were closely connected,and rich in extracellular matrix. Conclusion h PDLSC sheet was prepared successfully.