兔子宫平滑肌细胞的原代培养与鉴定  

Primary Culture and Identification of Rabbit Myometrial Smooth Muscle Cells

作　　者：曾润清[1] 刘庚贵[1] 刘惠萍[1] 李冰凌[1] 王若光[1] 

机构地区：[1]湖南中医药大学中西结合系,湖南长沙410007

出　　处：《实用预防医学》2007年第2期267-269,共3页Practical Preventive Medicine

基　　金：国家自然科学基金项目(30572405)

摘　　要：目的研究子宫平滑肌细胞(MSMC)的培养方法及一些生物学特性。方法运用酶消化后组织贴块法及胶原酶Ⅰ消化法进行兔子宫平滑肌细胞的培养,并用倒置生物显微镜、HE染色进行形态学观察和用免疫组化对培养细胞进行鉴定。结果运用此两种方法成功培养了兔子宫平滑肌细胞,培养的细胞呈典型“峰-谷”样生长,HE染色观察细胞呈梭形,胞浆丰富,核大圆形或椭圆形。免疫组化S-P法检测a-平滑肌肌动蛋白单克隆抗体(a-SMActin)表达强阳性。经传代纯化,纯度达99%以上,细胞生长特性未见异常改变。结论酶消化后组织贴块法及胶原酶Ⅰ消化法培养的兔子宫平滑肌细胞生长稳定性好,培养与纯化能同步进行,方法简便,经济。Objective To study the two methods of primary culture of rabbit myometrial smooth muscle cells （MSMC） and to identify their partial biological properties. Methods Rabbit myometrial tissue was planted and MSMC was digested by Collagenase I. The cellular morphology was observed by inverted phase contrast microscope and the nucleus and cytoplasm were observed after haematoxylin and eosin （HE） staining. The immunocytochemistry was used to identify the MSMC. Results The cells were successfully cultrured by these two methods and grown in the ＂peak-valley＂ mode. The cultrured MSMC were fusiform shape with a big oviform or round nuclei, and rich in cytoplasm. Immunohistochemical study revealed strong expression of a - smooth muscle actin （a--SMActin）, The purity of the MSMC was above 99 % after passage and purification, and no abnormality was seen. Conclusion We have successfully harvested the primary cultured rabbit myometrial smooth muscle cells and obtained monomorphic cells with stable growth. In addition, the method is convenient, economical and practical, and the culture and sublimation could be in synchronism.

关 键 词：兔 子宫平滑肌细胞 细胞培养 鉴定 原代培养 传代培养 

分 类 号：R-332[医药卫生] R339.22