摘要
旨在建立牦牛胚胎心脏原代成纤维细胞系并对其进行鉴定.以牦牛胚胎心脏组织块为研究对象,运用组织块法对其进行分离培养获得原代细胞,使用胰酶消化法对原代细胞进行传代培养,通过MTT法检测细胞增殖活性,利用HE染色法及秋水仙素处理染色体鉴定细胞外形,选取4个成纤维标志性基因TGF-β、FSP-1、S100A4、Vimentin,通过RT-qPCR技术对基因的表达情况进行鉴定.结果显示成功将原代分离培养的成纤维细胞传代至11代;通过外型的一系列检测方法发现其符合成纤维细胞的外形,该细胞活力充足,生长规律符合“S”型生长曲线;荧光定量检测以上四种成纤维细胞特征性基因在不同代数中均表达且相对稳定,结果证实分离培养出的细胞符合成纤维细胞系的分子表达特征.本研究成功建立了牦牛胚胎心脏原代成纤维细胞系,集成了该细胞系的鉴定流程,为今后牦牛进一步研究提供相关的细胞系平台.
The aim of this study was to establish and characterize a primary fibroblast cell line from yak embryonic heart.The primary cells were isolated and cultured using the tissue block method,and the cells were passaged by trypsin digestion.The cell proliferation activity was detected by MTT,and the cell shape was identified by HE staining and colchicine treatment of chromosomes.Four signature fibroblast genes,TGF-β,FSP-1,S100A4 and Vimentin,were selected and their expression was identified by RT-qPCR.The results showed that the fibroblasts isolated from the primary culture were successfully passed to 11 generations;the cells were found to be consistent with the appearance of fibroblasts by a series of external assays,and the cells were sufficiently vigorous and the growth pattern was in accordance with the“S”growth curve;the four fibroblast signature genes were expressed in different generations by fluorescence quantification and were relatively stable.The results confirmed that the isolated cells were consistent with the molecular expression characteristics of the synthetic fibroblast cell line.This study successfully established a primary fibroblast cell line from yak embryonic heart and integrated the identification process of this cell line,which will provide a relevant cell line platform for further research in yak.
作者
王玉玲
张露露
周珊
蒋涛
兰道亮
吉文汇
WANG Yu-ling;ZHANG Lu-lu;ZHOU Shan;JIANG Tao;LAN Dao-liang;JI Wen-hui(School of Animal Husbandry&Veterinary Sciences,Southwest Minzu University,Chengdu 610041,China;Ministry of Education Key Laboratory for Qinghai-Tibet Plateau Animal Genetic Resource and Utilization,Southwest Minzu University,Chengdu 610041,China)
出处
《西南民族大学学报（自然科学版）》
CAS
2022年第2期119-124,共6页
Journal of Southwest Minzu University(Natural Science Edition)
基金
中央引导地方科技发展专项(2020ZYD017)
西南民族大学中央高校基本科研业务费专项资金项目(2018NQN02)。
关键词
牦牛
心脏成纤维细胞
原代细胞系
分离培养
鉴定
yak
cardiac fibroblast
primary cell line
isolation and culture
identification
