离体培养人牙乳头细胞的消化法和贴壁法比较

Human dental papilla cells in culture ——comparision enzymatic separation with explants

目的：建立体外培养人牙乳头细胞的方法，比较酶消化法和组织块贴壁法对培养结果的影响。方法：分离合法堕胎的８月龄男胎的乳牙牙乳头组织，按Ａ、Ｃ区和Ｂ、Ｄ区将组织分成两组。一组用酶消化法获取细胞，即０．２％胰酶和０．０２％ＥＤＴＡ在３７℃下消化２０ｍｉｎ，另一组采用常规组织块贴壁法。结果：在酶消化法组中原代细胞按形态分为三种：梭形、星形纤维细胞样和内皮样。内皮细胞在贴壁培养组和继代的酶消化法组极少观察到。两种方法获得的细胞在传代和生长特性等方面相似。结论：酶消化法在来源组织丰富时是一种较好的原代培养方法，人牙乳头细胞在体外的成功培养将给人们研究该细胞提供一个重要的工具。

Aim: To establish an experimental model for culturing human dental papilla cells, and compare the effects of enzymatic separation and explant on culturing the human dental papilla cells. Methods: Dental papilla mesenchymes were obtained from an eight-month-old legally aborted male embryo. Before minced into less than 1 mm in diameter, mesenchymal tissues were divided into two groups according to A,C area and B,D area. The tissues of a random group were digested with 0.2% trypsin and 0.02% EDTA at 37℃ for 20 min. Fragments of the other group were explanted into flasks directly. Results: The papilla cells from two groups could grow progressively in DMEM medium containing 15%FCS.Primary culture cells were harvested more quickly from the group of enzymatic separation than those from the explant group. The primary cells from enzymatic group were morphologically divided into three types: spindle-shaped, stellate fibroblastic, and endothelial-like. The endothelial-like cells were hardly observed in primary culture of the other group and subculture of enzymatic group. Conclusions: It seemed that the enzymatic method was an efficient way to obtain the dental papilla cells in this study. The successful culture of human dental papilla cells now enables subsequent studies on the cellular properties related to the capability of differentiation and calcification of human papilla cells.