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.
文摘目的探讨CdCl_(2)与尼古丁联合作用对小鼠精原GC-1细胞周期和凋亡的影响及其联合作用方式。方法设置CdCl_(2)组、尼古丁组以及联合作用组24 h(简称联用组)3组,采用CCK-8法检测对GC-1细胞增殖的抑制,流式细胞术检测对GC-1细胞周期和凋亡的影响。结果随着浓度的升高,CdCl_(2)、尼古丁对GC-1细胞增殖的抑制作用增强,CdCl_(2)、尼古丁对GC-1细胞的24 h IC_(50)分别为5.409和2814μmol/L。CdCl_(2)和尼古丁联用,在尼古丁浓度为0.175、0.350、0.700、和1.400 mmol/L时,其对GC-1细胞的IC_(50)分别为4.422、4.532、3.309和2.532μmol/L,呈明显的递减过程,均低于CdCl_(2)单独作用组,CdCl_(2)和尼古丁联合作用时具有协同作用。细胞周期结果显示,CdCl_(2)浓度为2.5μmol/L时联用组G_(2)/M期细胞占比上升(P<0.01),与CdCl_(2)组、尼古丁组比较,联用组增强将细胞阻滞在G_(2)/M期的效应,对细胞周期的影响具有协同作用,且CdCl_(2)对于协同作用的影响更强。细胞凋亡结果显示,与CdCl_(2)组、尼古丁组比较,联用组细胞凋亡比例显著增加(P<0.01),对细胞凋亡的影响具有协同作用,且CdCl_(2)对于协同作用的影响更强。结论CdCl_(2)和尼古丁联合时具有协同作用,对小鼠精原细胞毒性作用增强,细胞周期发生阻滞,促进凋亡,且CdCl_(2)对协同作用的影响更强。
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.
