Coppe Carolyn, Zhang Yan, Den Besten Pamela K
Department of Oral Facial Sciences, Division of Pediatric Dentistry. University of California San Francisco, San Francisco, Calif, USA.
Pediatr Dent. 2009 Nov-Dec;31(7):467-71.
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.
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.
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.
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.
本研究旨在对人乳牙牙髓细胞进行特征分析，并确定其诱导口腔上皮细胞分化的能力。
从新鲜拔除的乳切牙中分离牙髓细胞，用4mg/ml胶原酶/ dispase消化，并在含有10%胎牛血清的杜氏改良 Eagle 培养基中培养。通过对 STRO-1 和 CD146 进行免疫细胞化学染色以及荧光激活细胞分选来鉴定干细胞群体。为了确定原代牙髓细胞是否能向上皮细胞发出信号，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。
对 STRO-1+细胞进行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%。与口腔上皮细胞共培养的生长停滞的原代牙髓细胞通过免疫细胞化学和PCR显示出釉原蛋白的表达。单独的口腔上皮细胞釉原蛋白免疫阴性。
乳牙牙髓细胞中干细胞含量不到2%。乳牙牙髓内的细胞可促进上皮细胞向成釉细胞表型分化，提示这种异质性细胞群体在细胞介导的釉质组织工程中有潜在应用价值。
