猪精原干细胞的原子力显微镜检测  

Observation of Pig Spermatogonial Stem Cell by Atomic Force Microscope

作　　者：李盛璞[1] 师如意[2] 王秋兰[1] 潘洁[1] 蔡继业[1] 张守全[2] 

机构地区：[1]暨南大学化学系,广东广州510632 [2]华南农业大学动物科学学院,广东广州510642

出　　处：《动物医学进展》2010年第6期30-34,共5页Progress In Veterinary Medicine

基　　金：国家973重大项目(2010CB833603);国家自然科学基金(60578025)

摘　　要：从幼年猪的睾丸中获取精原干细胞(SSC),并用原子力显微镜(AFM)在单细胞水平上分析SSC的形貌、超微结构和机械性能。试验中利用差速贴壁的方法分离纯化获得SSC,40 g/L的多聚甲醛固定SSC,空气中干燥后,在近生理条件下用AFM观察。由差速贴壁方法纯化后,SSC纯化率为91.61%±4.04%。从AFM的形貌图可知,SSC呈规则的圆形,中间隆起,超微结构显示SSC膜表面有一定的颗粒状物质存在,从附带的IP2.1软件可得出细胞的高低差Rp-v、均方根粗糙度Rq、平均粗糙度Ra、平均高度MeantHt等膜表面的几何参数。膜表面的黏弹力、硬度和杨氏模量等细微信息由AFM高空间分辨的力位移曲线测量系统得出。AFM对细胞的清晰成像和提供的表面定量信息为猪的SSC深入研究提供了可视化依据。The spermatogonial stem cells（SSC） from young pig testicular were obtained. The morphology, ultra-structure and mechanical property of pig SSC were analyzed by atomic force microscope （AFM） at single-cell level. Purification rate of SSC is 91.61%±4.04% by using differential adhesion method. The cells were fixed with 4 paraformal-dehyde and air-dried. The images showed that the shape of the cells presented round and there were a few of granular packed particles on the cell membrane. With the region analysis function of IP2.1 software, the regions of 1 /~m）〈 1 /Lm were selected and four parameters e. g. maximum peak-to-valley distance （Rp-v）, root-mean-squared roughness （Rq）, average roughness （Ra） and mean height （Meant Ht） of the cells were measured. From the high spatial resolution of AFM forcecurves, the mechanical properties （adhesion force, stiffness and Young＇s modulus） of the cells were obtained. It was concluded that AFM had specific advantages in analyzing cell membrane at nanometer level and could provide more precise information, which helped us increase the in-depth awareness of SSC.

关 键 词：猪精原干细胞 原子力显微镜 超微结构 

分 类 号：S852.11[农业科学—基础兽医学]