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目的:体外分离、培养山羊乳牙牙髓细胞,并转染绿色荧光蛋白。方法:采用组织块法获得山羊乳牙牙髓细胞,细胞计数法测定生长曲线,细胞爬片行HE染色、碱性磷酸酶染色、抗波形蛋白和角蛋白免疫组化染色。加入含GFP反转录病毒液转染山羊乳牙牙髓细胞。结果:通过组织块法获得了山羊乳牙牙髓细胞,细胞群体倍增时间为43.79h。HE染色显示细胞形态为梭形,细胞体小,胞核大。细胞碱性磷酸酶染色阳性,抗波形蛋白免疫组化染色阳性,抗角蛋白免疫组化染色阴性。转染绿色荧光蛋白后,细胞能持续表达绿色荧光。结论:山羊乳牙牙髓组织可分离培养出间叶来源的乳牙牙髓细胞,并成功使之表达绿色荧光。
OBJECTIVE: To isolate and culture goat deciduous dental pulp cells and transfect green fluorescent protein. Methods: Dendritic cells of goat milk decidua were obtained by tissue block method. The growth curve was determined by cell counting method. HE staining, alkaline phosphatase staining, anti-vimentin and keratin immunohistochemical staining were performed on the cells. Goat milk dental pulp cells transfected with GFP retrovirus were added. Results: Goat milk deciduous dental pulp cells were obtained by tissue block method. The doubling time of cell population was 43.79 hours. HE staining showed that the cell morphology was fusiform, small cell body and large nucleus. Alkaline phosphatase staining positive, anti-vimentin immunohistochemical staining positive, anti-keratin immunohistochemical staining was negative. After transfection with green fluorescent protein, cells can continuously express green fluorescence. Conclusion: The deciduous dental pulp cells derived from mesenchyme can be isolated and cultured from the pulp of goat deciduous teeth and the green fluorescent protein is successfully expressed.