猪牙乳头细胞的体外培养及生物学特性研究

目的:培养新生猪的牙乳头细胞,探讨其传代细胞的生物学特性。方法:选择刚出生1￣3d的新生猪,取出磨牙胚,分离牙乳头,用酶消化法培养猪牙乳头细胞,并用波形丝蛋白和细胞角蛋白鉴定细胞来源。观察细胞的生长规律,通过免疫组化染色检测Ⅰ型胶原、Ⅲ型胶原、牙本质涎蛋白(DSP)等的表达情况。结果:猪牙乳头细胞在体外培养时生长良好,波形丝蛋白染色阳性而细胞角蛋白染色阴性,细胞中Ⅰ型胶原、Ⅲ型胶原、DSP呈阳性表达。结论:通过酶消化法,可以体外扩增培养猪牙乳头细胞,细胞为间充质来源,其传代细胞在合成胶原及DSP的能力上与体内牙乳头细胞相似,提示猪牙乳头细胞可能具备作为牙髓-牙本质复合体再生研究种子细胞的潜能。

Culture and characteristics of porcine dental papilla cells (pDPCs) in vitro

PURPOSE: To culture porcine dental papilla cells (pDPCs) and to study the cytobiological characteristics of the cells in vitro. METHODS: Dental papillae were collected from postnatal (1-3 days) pigs, then the pDPCs were isolated and cultured through the method of enzyme-digestion. Vimentin and cytokeratin (CK) were used to demonstrate the cells' mesenchymal derivation. Collagen I collagen III and dentin sialoprotein (DSP) were detected in pDPCs by immunohistochemistry. RESULTS: The pDPCs were well cultured in the medium of DMEM/F12 containing 10% FBS in vitro. Vimentin was positively expressed in pDPCs but CK was not. Collagen I, collagen III, and DSP were all positive in the plasma of the pDPCs. CONCLUSIONS: The pDPCs were successfully isolated and cultured in vitro. The pDPCs come from mesenchymal tissue and have the capability of synthesizing extracellular matrix (collagen I, collagen III, and DSP) of dentin-pulp complex. This study indicated that the pDPCs may have the potential to be used as seed cell in regeneration of dentin-pulp complex.