福寿螺足组织和外套膜组织细胞培养的初步研究

采用改良的M199培养基,对福寿螺(Ampullaria gigas Spix)足组织和外套膜组织细胞进行了体外培养研究。结果显示:接种6 h后,细胞开始从组织块中迁出,培养至第3天,迁出细胞在组织块周围形成生长晕,第21~28天,细胞覆盖大部分培养瓶底壁。足组织和外套膜组织细胞分别被传至8代和9代。足组织和外套膜组织细胞在原代培养物中主要有两类,一为上皮样小细胞,形态为圆形、椭圆形或多边形,直径7~15μm;另一类为上皮样中型和大型细胞,形态椭圆形或多边形,直径15~30μm。上皮样小细胞数量多,增殖速度较快,在原代和传代培养的中后期皆可形成细胞单层。外套膜组织细胞的生长和增殖能力高于足组织细胞,其贴壁性也好于足组织细胞,Hoechst荧光染色显示外套膜组织细胞的细胞凋亡指数明显低于足组织细胞。结果表明,贝类外套膜组织有潜力成为建立连续细胞系的组织来源。

A Preliminary Study of Foot and Mantle Tissues Culture from Ampullaria gigas

The cells of foot and mantle tissues derived from Ampullaria gigas Spix were cultured in vitro with the modified medium 199.At 6 h post-inoculation,cells began to migrate out from the tissue.At 3 d post-inoculation the outgrowth was formed around the tissues.Being cultured for 21~28 days,most of the bottom of culture flask was covered by the cells.The cells of foot and mantle tissues have been subcultured to the eighth and ninth generation,respectively.There were two types of cells in the preliminary cell culture of the foot and mantle tissues.One was small epithelium-like cells,which were rounded,elliptic or polygonal with diameter of 7~15 μm;the other was middle epithelium-like size and large cells,which was elliptic and polygonal with diameter of 15~30 μm.There were plenty of the small epithelium-like cells,Which proliferated faster and could form cell monolayer in the preliminary culture and subculture.The results of preliminary culture and subculture showed that the growth ability and adherence of mantle tissue cells was better than that of foot tissue ones.The result of Hoechst fluorescein stain revealed that the apoptosis index of mantle tissue cells was apparently less than that of foot tissue cells.The present study implied that the mantle tissue could have potential to be established as a continuous cell line.