摘要
为建立崂山奶山羊骨髓间充质干细胞(BMSCs)永生化细胞系,将已构建的pcDNA3.1-EGFPTERT转入崂山奶山羊BMSCs中,利用含G418的培养基筛选获得稳定转染的TERT-BMSCs;通过多次传代,测定其生长曲线;选取高代次TERT-BMSCs的分裂中期相的细胞,检测其染色体及核型的稳定性。结果表明,转入TERT基因的崂山奶山羊BMSCs能够稳定表达该基因,其生长曲线呈"S"形;在多次传代后,P40代细胞的核型正常率仍能达到88.24%。综上提示,转染得到的TERT-BMSCs具有良好的生长状态,其细胞生长稳定,符合永生化细胞特征,为间充质干细胞应用于组织修复及基因工程种子细胞的制备提供了理论基础。
In order to establish immortalized bone marrow mesenchymal stem cells line of Laoshan dairy goat,a previously constructed plasmid vector pcDNA3.1-EGFP-TERT was transferred into bone marrow mesenchymal stem cells(BMSCs)of Laoshan dairy goat.The stable transfected TERT-BMSCs was screened by the culture medium containing G418.The growth curve of the obtained TERT-BMSCs was determined by multiple subcultures.The stable TERT-BMSCs cell line was established by determining the genetic stability of higher generation.The results showed that BMSCs of Laoshan dairy goat transferred by TERT gene was stable.The growth curve of TERT-BMSCs was S type.After multiple passages,the normal rate of nuclear type of the P40 TERT-BMSCs was 88.24%.In conclusion,the transfected TERT-BMSCs cells had a good growth state,and their growth was stable and consistent with the characteristics of immortalized cells.Our study provides a theoretical basis for the application of the mesenchymal stem cells(MSCs)in tissue repair and the preparation of genetic engineering seed cells.
作者
程明
刘开东
张宝旬
孙友德
李培培
杨培培
王建华
刘宗正
CHENG Ming;LIU Kai-dong;ZHANG Bao-xun;SUN You-de;LI Pei-pei;YANG Pei-pei;WANG Jian-hua;LIU Zong-zheng(Qingdao Animal Husbandry and Veterinary Medicine Research Institute,Qingdao 266100,China;Qingdao Aote Breeding Sheep Farm,Qingdao 266100,China)
出处
《畜牧与饲料科学》
2019年第1期11-14,共4页
Animal Husbandry and Feed Science
基金
农业农村部华东动物营养与饲料科学观测实验站运行经费
青岛市财政专项资金项目
