摘要
目的 摸索正常胎鼠牙胚细胞离体培养所需条件 ,为进一步将其作为牙组织工程种子细胞奠定基础。方法 体视显微镜下取 17d龄胎鼠下颌第一磨牙牙胚 ,胶原酶 /分离酶消化 ,以含 10 %胎牛血清 (FCS)的DMEM为培养基进行原代培养。倒置显微镜下观察培养细胞的生长特点 ,MTT法测定细胞活性绘制生长曲线 ,取原代培养 4～ 9d的细胞进行组织化学和免疫组织化学染色。结果 倒置显微镜下观察 ,培养细胞在 2 4h内贴壁 ,约 7～ 9d生长连成片状 ,细胞有多角形、大而扁平梭形和长梭形几种形态。生长曲线表明 :培养细胞在前 3天生长缓慢 ,第 4天呈对数生长 ,第 7天达高峰。免疫组化结果表明 :培养细胞来源于两种组织。结论 采用酶消化培养法 ,用胶原做基质饲养层 ,含 10 %FCS的低糖DMEM作为培养基 ,能将胎鼠牙胚细胞培养成活 ,并维持其细胞学特征。
Objective To explore the essential conditions of the normal tooth germ cells culture in order to use these cultured cells as the seeding cell in the tissue-engineering of tooth .Methods Take out the first mandibular molar tooth germs carefully from 17 days fetal SD rats in use of dissectal anatomic micrograph. Enamel and pulp organ tissues were enzymatically treated into single cell and cell units suspension and cultured in Low Glucose DMEM with 10% Fetal Cow Serum(FCS). Observe the cells growth characteristics on Covert Micrograph, and draw a growth curve after MTT essay. Identify these cells by morphology and immunohistochemistry . Results Cells adhered to the plates within 24 hours and growth in monolayer before confluence.Cells cultured in DMEM formed a mixed cell population of epithelial-like and fibroblast-like.The multilateral-shaped cells grow tightly in typical cobblestone appearance. The growth curve indicate that cells proliferate slowly in the first 3 days, then increased gradually. The two generation cells of tooth germ can be cultured in one medium by immunohistochemistry. Conclusions By enzymatically treated, collagen used as substrate, tooth germ cells could be mixed cultured and maintained its characteristics.
出处
《锦州医学院学报》
2004年第4期24-28,共5页
Journal of Jinzhou Medical College
关键词
牙胚细胞
培养
胎鼠
牙组织工程
tooth germ cell
cultured
fetal rats
tissue-engineering of tooth
