Characterization of primary dental pulp cells in vitro.
Characterization of primary dental pulp cells in vitro.
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DOI：
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发表时间：
2009-11
影响因子：
1.6
通讯作者：
C. Coppe;Yan Zhang;P. Den Besten
中科院分区：
文献类型：
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作者：
C. Coppe;Yan Zhang;P. Den Besten
目的研究人乳牙牙髓细胞的特性及其诱导口腔上皮细胞分化的能力。方法从新鲜拔除的乳牙中分离牙髓细胞，用4 mg/ml胶原酶/分散酶消化，在含10%胎牛血清的Dulbecco改良Eagle培养基中培养。通过STRO-1和CD 146免疫细胞化学染色和荧光激活细胞分选鉴定干细胞群。为了确定原代牙髓细胞是否可以向上皮发出信号，将牙髓细胞与人胎儿口腔上皮细胞共培养。3天后，收集共培养的细胞，并通过聚合酶链反应（PCR）和免疫细胞化学染色分析釉原蛋白的表达。结果STRO-1+细胞的免疫荧光和荧光激活细胞分选显示该干细胞群体约占总群体的2%。免疫细胞化学和PCR检测显示，与口腔上皮细胞共培养的生长停滞的原代牙髓细胞表达釉原蛋白。单独的口腔上皮细胞为釉原蛋白免疫阴性。结论：乳牙牙髓细胞中干细胞含量不足2%。乳牙牙髓内的细胞可以促进上皮细胞向成釉细胞表型分化，这表明这种异质细胞群在细胞介导的釉质组织工程中的潜在用途。
PURPOSE This study's purpose was to characterize dental pulp cells from human primary teeth and determine their ability to induce differentiation of oral epithelial cells. METHODS Dental pulp cells were isolated from freshly extracted primary incisors, digested with 4 mg/ml collogenase/dispase, and grown in Dulbecco's modified Eagle's medium with 10 percent fetal bovine serum. Stem cell populations were identified by immunocytochemical staining for STRO-1 and CD146 and fluorescence activated cell sorting. To determine whether primary pulp cells can signal epithelium, the pulp cells were grown in coculture with human fetal oral epithelial cells. After 3 days, the cocultured cells were collected and analyzed for amelogenin expression by polymerAse chain reaction (PCR) and immunocytochemical staining. RESULTS Immunofluorescence and fluorescence activated cell sorting of STRO-1+ cells showed this stem cell population to be approximately 2 percent of the total population. Growth-arrested primary dental pulp cells grown in coculture with oral epithelial cells showed expression of Amelogenin by immunocytochemistry and PCR. Oral epithelial cells alone were amelogenin immunonegative. CONCLUSIONS Primary tooth dental pulp cells contain less than 2 percent stem cells. Cells within the primary tooth pulp can promote epithelial cell differentiation toward an ameloblast phenotype, suggesting the potential use of this heterogeneous population of cells in cell-mediated enamel tissue engineering.