医学论文-大鼠牙囊细胞培养方法的探讨.docVIP

医学论文-大鼠牙囊细胞培养方法的探讨 【摘要】? 【目的】 探讨大鼠牙囊细胞体外培养的方法。【方法】 分离出大鼠牙囊组织，分别采用组织块培养法、细胞消化分散法、及改良组织块法进行牙囊细胞原代及传代培养，倒置相差显微镜下进行细胞形态学观察，透射电镜观察细胞超微结构。【结果】 培养至第3天，倒置相差显微镜观察发现，采用组织块培养法，见少数细胞从组织块爬出；而细胞消化分散法则可获得数量多的牙囊细胞，但与杂质细胞混杂生长；改良组织块法见量多、相对纯的牙囊细胞。电镜下牙囊细胞无桥粒，胞浆中含有高密度电子颗粒和大量粗面内质网。组织块法、细胞消化分散法和改良组织块法牙囊细胞培养成功率分别为80%、60%和100%；分别于10 d、7 d和6 d左右进行第一次传代。 【结论】 改良组织块法是一种良好的大鼠牙囊细胞培养方法。 【关键词】? 牙囊细胞； 细胞培养； 方法学 ??? Abstract： 【Objective】 To explore the culture method of rat dental follicle cells in vitro. 【Methods】 The dental follicle (DF) tissues were dissected for primary generation and passage culture by methods of explant, dissociation cell and improved explant, respectively. Consequently, the morphology of the cultured first passage DF cells and their ultrastructure were discovered under inversion phase difference microscope and transmission electronic microscope, respectively. 【Results】 At the third day in vitro, only a few cells extended from the explant by way of explant, and many DF cells mixing with impure cells adhered wall by way of dissociation cell. However, a large quantity of relative pure DF cells was discovered by way of improved explant at the same time. Transmission electron microscopy showed? that the cultured cells contained electron-dense granules, and abundant rough endoplasmic reticulum（RER） and no desmosmoes. At about 10th , 7th and 6th day in vitro, DF cells were passed first time with achievement ratio of eighty, sixty and one hundred percent by ways of explant, dissociation cell? and improved explant culture respectively. 【Conclusion】 Improved explant culture was a good culturing method for rat DF cells. ??? Key words: dental follicle cells； cell culture； methodology ??? 牙囊对牙齿萌出起重要作用。牙萌出必须是在牙槽骨的吸收、形成牙槽骨的通道后，才能够完成这一生理过程[1]。Cahill和Marks[2，3]研究表明，以手术方法去除小鼠牙囊，牙齿则不能萌出；而保留牙囊，用其它物质替代牙胚，结果替代物可以萌出。Larson[4]在狗前磨牙牙冠形成后萌出前四周去除牙囊，牙齿同样不能萌出，将剥离的牙囊立即放回成釉器表面，牙齿即能重新萌出。这些研究证明牙齿萌出依赖牙囊的存在。牙囊在牙齿萌出过程中，从组织、细胞和分子水平上对牙齿萌出起重要的调控作用[5，6]。目前关于牙囊细胞的体外培养，国内外报道尚不多[7]。本研究以SD大鼠为实验对象，采用组织块培养法、细胞消化分散法及改良组织