【摘要】 目的:通过优化条件培养基用于非牙源性上皮细胞的体外扩增,为进一步开展组织工程牙提供上皮种子细胞来源奠定基础。方法:获取出生后1 d C57BL鼠原代口腔黏膜上皮细胞,接种于含有Y27632的上皮条件培养基的培养瓶内。细胞长满后,消化传代,采用条件培养基与3T3饲养层细胞共培养进行扩增并连续传代,观察细胞生长情况及连续传代后的生物学特性,免疫组织化学进行干细胞鉴定。结果:与传统上皮细胞培养方式相比,含Y27632的条件性培养基加3T3饲养层方式培养的上皮细胞生长迅速,细胞呈立方形或多角形,呈铺路石样排列。免疫组织化学染色表明这些干细胞表达CK14,SOX2和LGR5。连续传代至第7代后,细胞形态、生长曲线和干细胞标志没有明显变化。结论:含有Y27632的条件培养基可有效用于体外大量扩增非牙源性上皮细胞,并能保持干细胞生长特性,有望用于组织工程牙的种子细胞来源。更多还原

【Abstract】 Objective: To characterize the role of conditioned culture media on expansion of non-odontogenic epithelial cells in vitro for potential seed cells of further tissue engineering tooth. Methods: The oral mucosa epithelial cells were harvested from 1day postnatal mouse and cultured to confluency in conditioned media supplemented with Y27632,then passaged and plated onto 3T3 feeder cells in conditioned media with Y27632 for continuous passage. Histology and immunohistochemistry were performed for biological characteristics analysis of cells. Results: Compared with the traditional ways of epithelial cell culture,the oral mucosa epithelial cells plated onto3T3 feeder cells in conditioned media supplemented with Y27632 grew rapidly,which showed a cuboidal,polygonal or pavement-like shape and expressed CK14,SOX2 and LGR5. After continuous passage,the cell morphology,growth curve and cell markers have no obvious changes. Conclusions: The conditioned culture media supplemented with Y27632 have potential to effectively expand non-odontogenic epithelial cells in vitro,and maintain the characteristics of stem cells,which is expected for tissue engineering tooth as seed cells.更多还原