一株高分化效率的永生化猪脂肪前体细胞系建立  

Establishment of an immortalized porcine subcutaneous preadipocytes cell line with high differentiation efficiency

作　　者：洪鹏程 程运谋 朱海宁 余倩 黄赞 HONG Pengcheng;CHENG Yunmou;ZHU Haining;YU Qian;HUANG Zan(National Center for International Research on Animal Gut Nutrition/Laboratory of Gastrointestinal Microbiology,Nanjing Agricultured University,Nanjing 210095,China)

机构地区：[1]南京农业大学国家动物消化道营养国际联合研究中心/消化道微生物实验室,江苏南京210095

出　　处：《南京农业大学学报》2023年第1期128-134,共7页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(32170847,31500944)。

摘　　要：[目的]本研究旨在获得可以稳定传代并且始终保持高分化效率的猪脂肪前体细胞系。[方法]采集1日龄约克夏仔猪背部皮下脂肪组织,分离原代猪皮下脂肪前体细胞(porcine subcutaneous preadipocytes,PSPA),利用过表达SV40T的慢病毒感染从而获得永生化的PSPA,进一步通过有限稀释法获得了107株单克隆细胞系,从中筛选出一株具有高分化效率的单克隆细胞系,命名为Pig#4。通过比较相近代数时Pig#4与永生化PSPA、3T3-L1小鼠脂肪前体细胞系的分化效率,验证单克隆细胞在高代数时的分化效率。[结果]Pig#4 PSPA细胞在高传代数(P30)仍具有与永生化的PSPA细胞系相似的外形特征和增殖能力,仍能保持与3T3-L1细胞系近似的分化效率。并且,相较于原代PSPA细胞,Pig#4 PSPA细胞具有更高表达分化与聚脂成熟相关基因的趋势。[结论]获得了1株可以稳定传代并保持高分化效率的永生化猪脂肪前体细胞系。[Objectives]This study aimed to establish an immortal porcine preadipocytes cell line with high adipogenesis efficiency. [Methods]The primary porcine subcutaneous preadipocytes(PSPA)were obtained from the back fat of a 1-day-old Yorkshire piglet and immortalized by SV40T-overexpress-lentivirus. From the immortalized PSPA,107 monoclonal cell lines were picked with the limit dilution method. The monoclonal cell line with the highest adipogenesis efficiency was screened out and named as Pig#4. The adipogenesis efficiency of Pig#4,immortalized PSPA,and 3T3-L1 mouse preadipocytes cell lines were compared, we showed that Pig#4 had the highest adipogenesis efficiency, even in high passage. [Results]The morphological characteristics of Pig#4 in high passage(P30)were similar to immortalized PSPA,and the adipogenesis efficiency was comparable to 3T3-L1. Moreover, compared with the primary PSPA cells, Pig#4 PSPA cells had a higher expression of differentiation and adipogenic related genes. [Conclusions]We established an immortalized porcine subcutaneous preadipocytes cell line with high adipogenesis efficiency.

关 键 词：猪脂肪前体细胞 高分化效率 细胞系 诱导分化 

分 类 号：S828.9[农业科学—畜牧学]