Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue cul...Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue culture（MethodⅢ）,and cultural behaviors of cells were observed.The results showed that typical spermatogonium colonies appeard at 144 h of culture by enzymatic digestion-percoll density gradient centrifugation method and tubular fragments culture method,2.5%FBS kept the characteristics of spermatogonium stem cell better than others,produced more mass clones,and FBS of more than 2.5%concentration benefited spermatogonium differentiation and the number of colonies was significantly affected by FBS concentration.After 1 week of culture in method Ⅲ,the diameter of lumens and quantity of sertoli’s cells in tubal wall increased obviously,lumen of seminiferous tubules appeared.Sertoli’s cells kept constant and the number of spermatogoniums decreased obviously after 2 weeks of culture.展开更多
中华绒螯蟹(Eriocheir sinensis H.Milne Edwards)属甲壳纲(Crustacea)、软甲亚纲(Malacostraca)、十足目(Decapoda)、方蟹科(Grapsidae),俗称河蟹,是我国重要的经济蟹类,其形态解剖、生态习性及生理等方面的研究已有报道(Panning,1939;...中华绒螯蟹(Eriocheir sinensis H.Milne Edwards)属甲壳纲(Crustacea)、软甲亚纲(Malacostraca)、十足目(Decapoda)、方蟹科(Grapsidae),俗称河蟹,是我国重要的经济蟹类,其形态解剖、生态习性及生理等方面的研究已有报道(Panning,1939;Koch,1952;堵南山,1954,1957,1958;Bauchau,1960;Leersnyder,1966,1967;Dbainaut et al.,1976;Chevigue,1976;Leeranyder et al.,1977,1978;Pequenx et al.,1982;Chapelle et al.,1982;谈奇坤等,1984.).展开更多
目的:探讨大鼠精原细胞的分离纯化. 方法:采用酶消化法制备10 d Wister大鼠睾丸单细胞悬液;Percoll液不连续密度梯度分离精原细胞;用贴壁速度的差异纯化精原细胞.光电镜观察精原细胞的形态结构特征;应用酪氨酸蛋白激酶(c-kit)免疫化学...目的:探讨大鼠精原细胞的分离纯化. 方法:采用酶消化法制备10 d Wister大鼠睾丸单细胞悬液;Percoll液不连续密度梯度分离精原细胞;用贴壁速度的差异纯化精原细胞.光电镜观察精原细胞的形态结构特征;应用酪氨酸蛋白激酶(c-kit)免疫化学鉴定精原细胞. 结果:精原细胞主要分布在28%～36%(Ⅱ带)间的Percoll梯度;分离纯化的精原细胞形态结构与组织切片上精原细胞一致,其纯度达55.68%;c-kit在精原细胞呈阳性表达,在睾丸体细胞呈弱阳性或阴性表达.结论:利用酶消化法、Percoll液不连续密度梯度及细胞贴壁速度的差异,分离纯化的精原细胞满足体外实验要求;精原细胞表达特异的c-kit受体.展开更多
基金Supported by Fund of Heilongjiang Acadamy of Agricultural Science
文摘Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue culture（MethodⅢ）,and cultural behaviors of cells were observed.The results showed that typical spermatogonium colonies appeard at 144 h of culture by enzymatic digestion-percoll density gradient centrifugation method and tubular fragments culture method,2.5%FBS kept the characteristics of spermatogonium stem cell better than others,produced more mass clones,and FBS of more than 2.5%concentration benefited spermatogonium differentiation and the number of colonies was significantly affected by FBS concentration.After 1 week of culture in method Ⅲ,the diameter of lumens and quantity of sertoli’s cells in tubal wall increased obviously,lumen of seminiferous tubules appeared.Sertoli’s cells kept constant and the number of spermatogoniums decreased obviously after 2 weeks of culture.
文摘中华绒螯蟹(Eriocheir sinensis H.Milne Edwards)属甲壳纲(Crustacea)、软甲亚纲(Malacostraca)、十足目(Decapoda)、方蟹科(Grapsidae),俗称河蟹,是我国重要的经济蟹类,其形态解剖、生态习性及生理等方面的研究已有报道(Panning,1939;Koch,1952;堵南山,1954,1957,1958;Bauchau,1960;Leersnyder,1966,1967;Dbainaut et al.,1976;Chevigue,1976;Leeranyder et al.,1977,1978;Pequenx et al.,1982;Chapelle et al.,1982;谈奇坤等,1984.).
文摘目的:探讨大鼠精原细胞的分离纯化. 方法:采用酶消化法制备10 d Wister大鼠睾丸单细胞悬液;Percoll液不连续密度梯度分离精原细胞;用贴壁速度的差异纯化精原细胞.光电镜观察精原细胞的形态结构特征;应用酪氨酸蛋白激酶(c-kit)免疫化学鉴定精原细胞. 结果:精原细胞主要分布在28%～36%(Ⅱ带)间的Percoll梯度;分离纯化的精原细胞形态结构与组织切片上精原细胞一致,其纯度达55.68%;c-kit在精原细胞呈阳性表达,在睾丸体细胞呈弱阳性或阴性表达.结论:利用酶消化法、Percoll液不连续密度梯度及细胞贴壁速度的差异,分离纯化的精原细胞满足体外实验要求;精原细胞表达特异的c-kit受体.
