Analysis of developmental potentials of dental pulp in vitro using GFP transgenes.
Analysis of developmental potentials of dental pulp in vitro using GFP transgenes.
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DOI：
10.1111/j.1601-6343.2005.00347.x
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发表时间：
2005-11-01
影响因子：
3.1
通讯作者：
Mina, M
中科院分区：
文献类型：
--
作者：
Balic, A;Mina, M
背景：近年来，从成人组织中鉴定干细胞及其在牙齿替换/再生中的潜在应用方面取得了越来越多的进展。我们之前的体内研究表明，pOBCol3.6GFP和pOBCol2.3GFP转基因动物为深入了解牙髓中能够分化为成牙本质细胞的前体/干细胞提供了一个独特的模型。目的：研究pOBCol3.6GFP动物来源的牙髓细胞在体外的行为特征。实验设计：从出生5天的pOBCol3.6GFP杂合小鼠的第一磨牙牙髓分离出原代培养物，培养21天。结果：我们的观察结果表明，3.6-GFP来源的牙髓细胞含有大量具有矿化能力的增殖克隆细胞。我们还显示了3.6-GFP在牙髓原代培养中的阶段特异性激活/上调。在这些培养物中，Col1a1-3.6-GFP的表达发生在矿化结节出现之前，并且在矿化结节中不受调控。结论：Col1a1-GFP转基因似乎满足了完整牙齿和牙髓原代培养细胞谱系研究的许多标记基因的要求。
BACKGROUND: In recent years there has been increasing progress in identifying stem cells from adult tissues and their potential application for tooth replacement/regeneration. Our previous in vivo studies show that pOBCol3.6GFP and pOBCol2.3GFP transgenic animals provide a unique model to gain insight into progenitor/stem cells in the dental pulp capable of giving rise to odontoblasts.OBJECTIVES: To characterize the behavior of dental pulp cells derived from pOBCol3.6GFP animals in vitro.EXPERIMENTAL DESIGN: Primary cultures were established from the coronal portions of the pulps isolated first molars from 5-day-old pOBCol3.6GFP heterozygous mice and grown for 21 days. In these cultures proliferation, clonogenic capacity, activation of 3.6-GFP and mineralization were examined.RESULTS: Our observations show that dental pulp cells derived from 3.6-GFP contain a population of proliferative, clonogenic cells with the ability to mineralize. We also show the stage specific activation/upregulation of 3.6-GFP in primary cultures derived from dental pulp. In these cultures, expression of Col1a1-3.6-GFP occurs prior to the appearance of mineralized nodules and is unregulated in mineralized nodules.CONCLUSIONS: Col1a1-GFP transgenes appear to fulfill many of the requirements of a marker gene for cell lineage studies in intact tooth and primary cultures derived from dental pulp.