版纳微型猪近交系新生猪成纤维细胞的体外培养及其生物学特征研究  

In Vitro Culture and Biological Characteristics of Piglet Fibroblast Cell in Banna Mini-pig Inbred Line

作　　者：许成盛[1] 魏红江[1] 李红[1] 汪霞[2] 杨朝霞 卿玉波[1] 潘伟荣[1] 信吉阁[1] 曾养志[1] 

机构地区：[1]云南农业大学,云南省版纳微型猪近交系重点实验室,云南昆明650201 [2]云南农业大学动物科学技术学院,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2011年第3期323-327,共5页Journal of Yunnan Agricultural University:Natural Science

基　　金：云南省教育厅科研基金(07Z11542)

摘　　要：试验以版纳微型猪近交系新生猪耳部皮肤组织为材料,采用胰蛋白酶消化法培养成纤维细胞,并对其进行细胞形态观察、细胞冻存前和复苏后存活率测定、生长曲线绘制和染色体核型分析等生物学特征检测。结果表明:版纳微型猪近交系新生猪成纤维细胞呈典型的成纤维细胞形态;细胞冻存前和复苏后存活率分别为98.0%和94.07%;生长曲线呈"S"型,倍增时间为33 h;对所制备的染色体核型分析显示2n=38(XY),并在体外培养14代后仍能保持正常核型。本研究建立了稳定的版纳微型猪近交系新生猪成纤维细胞的培养方法,将为体细胞克隆、转基因和异种器官移植等相关的研究提供技术基础。In this study, the fibroblast cell which isolated from a newborn piglet ear skin of Banna mini-pig inbred line was used as the study materials to establish a fibroblast cell line by using the cold trypsinization method. The present research was designed to examine the characteristics of piglet fibroblast which involved the observations of cell morphology, determination of the cell viability before freezing and after thawing, dynamic growth curve and the chromosome nuclear type analysis. The results showed that the cells was morphologically consistent with fibroblasts shaped, cell viabilities before freezing and after thawing were 98.00% and 94. 07%, respectively. Additionally, the dynamic growth curve showing a sigmoid curve with population doubling time （PDT） for cells growth was approximately 33 h, the karyotype result gave a 2n = 38 （XY） and the karyotype was normal after cultured up to passage 14 in vitro. In conclusion, the present study has successfully established the newborn piglet ear fibroblast cell line in Banna mini-pig. It not only can preserve the resource on the cell level but also can be used as donor karyoplast for somatic cell nuclear transfer; produce transgenie animals, and xenotransplantation as well as related research in the future.

关 键 词：版纳微型猪近交系 成纤维细胞 细胞培养 生物学特征 

分 类 号：S828.91[农业科学—畜牧学]