摘要
背景:牙髓干细胞的多分化潜能、高扩增速率和容易获取使之成为引人注目的间充质干细胞来源。目的:观察兔牙髓细胞增殖特性并进行细胞表面标志物的鉴定。方法:体外分离培养兔牙髓细胞,采用酶解组织块法培养细胞至第3代观察细胞形态变化、计数细胞活率、细胞克隆形成率、细胞生长曲线、细胞增殖周期以及细胞表面标志物的鉴定。结果与结论:酶解组织块法可以较快的收获各代兔牙髓细胞。第3代到第6代细胞活率分别比为94.7%∶95.8%∶95.2%∶95.3%,细胞的克隆形成率为18个/2000;细胞的生长曲线基本符合间充质细胞特征,细胞周期G0/G1期大于80%;免疫细胞化学vimentin、CD44、osteonectin、Dsp均为阳性表达。证实兔牙髓细胞中可分离培养出牙髓干细胞,并能在体外有效增殖。
BACKGROUND： The multi-lineage differentiation potential, high amplification rate and accessibility of the dental pulp cells make them to become an attractive source of mesenchymal stem cells. OBJECTIVE： To observe the proliferation characteristics of rabbit dental pulp cells and to identify the cells surface marker METHODS： The dental pulp cells were isolated and cultured in vitro, then the cells were cultured to the third generation by enzymatic digestion method to observe the morphological changes, calculate the cell survival rate, test the cell clone forming rate, measure the cell growth curve and the cell proliferation cycle, and identify the cell surface markers. RESULTS AND CONCLUSION： The enzymatic digestion method could rapidly harvest various generations of rabbit dental pulp cells. The proportion of the cell survival rate was 94.7%：95.8%：95.2%：95.3% from the third generation to the sixth generation. The cells clone forming rate was 18/2 000; the cell growth curve was in line with the characteristics of mesenchymal cells, and the cell cycle rate of Go and G1 was more than 80%; cell multiplication cycle DNA purity more than 80 %. Immunocytochemistry staining showed the positive expression of vimentin, CD44, osteonectin and dentin sialoprotein. It indicates that the dental pulp stem cells can be isolated from the rabbit dental pulp cells and effectively proliferated in vitro.
出处
《中国组织工程研究》
CAS
CSCD
2012年第49期9236-9240,共5页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(30960423)~~
