动物营养学报
         首页        期刊介绍        编委会         编辑部         投稿须知        英文刊IFA        会议信息        联系我们      留言与回复
动物营养学报 2018, Vol. 30 Issue (9) :3551-3558    DOI: 10.3969/j.issn.1006-267x.2018.09.025
反刍动物营养 Ruminant nutrition 最新目录 | 下期目录 | 过刊浏览 | 高级检索 << Previous Articles | Next Articles >>
绒山羊羔羊和成年羊前体脂肪细胞的原代培养及传代方法
张清月, 王雪, 刘树林, 于洋, 郭晓宇, 闫素梅
内蒙古农业大学动物科学学院, 呼和浩特 010018
Primary Culture and Passage Method of Preadipocytes of Cashmere Lambs and Adult Goats
ZHANG Qingyue, WANG Xue, LIU Shulin, YU Yang, GUO Xiaoyu, YAN Sumei
Collage of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China
Download: PDF (6800KB)   HTML (1KB)   Export: BibTeX or EndNote (RIS)      Supporting Info
摘要 本试验主要探索绒山羊羔羊和成年羊前体脂肪细胞的培养及传代方法,为研究绒山羊的脂肪代谢机理提供细胞模型。以3月龄阿尔巴斯白绒山羊羔羊肾周脂肪组织为试验材料,采用胶原酶法直接得到羔羊前体脂肪细胞,结合其细胞形态观察、生长曲线和油红O染色进行鉴定;以绒山羊成年羊肾周脂肪组织为试验材料,采用胶原酶法和"天花板"法得到成熟脂肪细胞后,通过去分化得到前体脂肪细胞,诱导分化后利用油红O染色鉴定其向成熟脂肪细胞分化的情况。绒山羊羔羊肾周脂肪组织中前体脂肪细胞的适宜分离条件为0.1%Ⅰ型胶原酶、37℃消化1 h,250×g离心10 min;传代时分离细胞采用0.25%胰蛋白酶消化60 s。细胞形态为梭形,生长曲线呈"S"型,油红O染色结果呈阳性。成年羊采用和羔羊相同的胶原酶法获得成熟脂肪细胞后,通过改进的"天花板"法获得前体脂肪细胞,经诱导分化后,油红O染色结果呈阳性。综上,采用Ⅰ型胶原酶消化法可直接分离培养3月龄绒山羊羔羊的前体脂肪细胞;采用胶原酶法和改进后的"天花板"法分离培养成年绒山羊的前体脂肪细胞是可行的。
Service
把本文推荐给朋友
加入我的书架
加入引用管理器
Email Alert
RSS
作者相关文章
关键词绒山羊成年羊   绒山羊羔羊   前体脂肪细胞   细胞培养     
Abstract: The study aimed to explore the primary culture and passage methods of preadipocytes of cashmere lambs and adult goats, and to provide a cell model for further researching the metabolic mechanism of adipocytes of cashmere goats. Perirenal adipose tissues of 3-month-old Albas cashmere lambs were selected as test materials. Preadipocytes of lambs were obtained directly by collagenase digestion method, and identified by morphological observation, growth curve and oil red O staining; perirenal adipose tissues of cashmere adult goats were selected as test materials. Mature adipocytes of adult goats were obtained by collagenase digestion and "ceiling" culture methods, and then preadipocytes were obtained by the dedifferentiation of mature adipocytes. After induction of differentiation, oil red O staining was used to identify their differentiation into mature adipocytes. The appropriate separation conditions for preadipocytes of the perirenal adipose tissues of lambs were digestion by 0.1% Ⅰ collagenase at 37℃ for 1 h, and centrifugation at 250×g for 10 min; during cell passage, digestion by 0.25% trypsin for 60 s. The preadipocytes were fusiform, grew in an "S" shape, and were positive on oil red O staining. Mature adipocytes of adult goats were obtained by collagenase digestion method in the same way as the lambs' method, and then using modified "ceiling" method obtained preadipocytes. After induction of differentiation, those preadipocytes were positive on oil red O staining. In conclusion, it is feasible to obtain preadipocytes directly by type Ⅰ collagenase digestion method from 3-month-old Albas cashmere lambs and by collagenase digestion and modified "ceiling" methods from adult cashmere goats.
Keywordscashmere adult goats,   cashmere lames,   preadipocytes,   cell culture     
收稿日期: 2018-02-27;
基金资助:

国家自然科学基金(31760685)

通讯作者 闫素梅,教授,博士生导师,E-mail:yansmimau@163.com     Email: yansmimau@163.com
作者简介: 张清月(1993-),女,河南新乡人,硕士研究生,从事反刍动物营养的研究。E-mail:alicezqy@126.com
引用本文:   
. 绒山羊羔羊和成年羊前体脂肪细胞的原代培养及传代方法[J]. 动物营养学报, 2018,V30(9): 3551-3558
. Primary Culture and Passage Method of Preadipocytes of Cashmere Lambs and Adult Goats[J]. Chinese Journal of Animal Nutrition, 2018,V30(9): 3551-3558.
链接本文:  
http://www.chinajan.com/CN/10.3969/j.issn.1006-267x.2018.09.025     或     http://www.chinajan.com/CN/Y2018/V30/I9/3551
 
[1] 郭红芳,昝林森,孙永刚.牛前体脂肪细胞的分离培养及诱导分化[J].西北农林科技大学学报,2014,42(2):1-6,12.
[2] 蔡勇,阿依木古丽,杨具田,等.绵羊前体脂肪细胞的原代培养及分化[J].动物营养学报,2010,22(6):1768-1774.
[3] 李影,杨公社,卢荣华,等.原代猪前体脂肪细胞培养方法的优化[J].细胞生物学杂志,2005,27(6):697-700.
[4] 杜琛,付绍印,韩志玲,等.绒山羊肌内前体脂肪细胞的基因表达分析[J].畜牧兽医学报,2013,44(10):1532-1538.
[5] 李翼飞,王兆杰.脂肪细胞去分化过程中脂滴丢失的调控机制研究进展[J].西部医学,2015,27(8):1271-1274.
[6] 何衡杰,郝丽.肿瘤坏死因子α在超重者脂肪组织中的表达[J].中国卫生产业,2014,23(11):13-14,17.
[7] 陈晓炜,姜平,高建华,等.脂肪细胞去分化及构建组织工程化脂肪的实验研究[J].南方医科大学学报,2009,29(4):606-610.
[8] 王欢欢,孙文星,徐春瑛,等.猪成熟脂肪细胞分离培养及去分化研究[J].农业生物技术学报,2012,20(8):915-921.
[9] 吴迪.去分化脂肪细胞与脂肪干细胞成骨及成软骨能力的比较研究[D].硕士学位论文.遵义:遵义医学院,2012.
[10] 宋子仪,史新娥,杨浩,等.基于一种新的天花板培养方法分析猪成熟脂肪细胞去分化过程中关键基因表达模式[J].农业生物技术学报,2013,21(4):379-387.
[11] 司徒镇强,吴军正.细胞培养[M].2版.西安:世界图书出版公司,2007.
[12] 胡艳霞,曾勇庆,崔志峰,等.莱芜猪前体脂肪细胞的原代培养及诱导分化研究[J].畜牧兽医学报,2012,43(9):1346-1352.
[13] 孙超.ECM组分和cAMP对大鼠前体脂肪细胞增殖与分化的调控[D].博士学位论文.杨凌:西北农林科技大学,2001.
[14] 屈长青,张国华,陈粉粉,等.猪前体脂肪细胞的原代培养[J].农业生物技术学报,2005,13(5):649-653.
[15] NG C G,POZNANSKI W J,BOROWIECKI M,et al.Differences in growth in vitro of adipose cells from normal and obese patients[J].Nature,1971,231(5303):445.
[16] 张艳芳,任阳,朱琳娜,等.猪肌内前体脂肪细胞的体外培养[J].农业生物技术学报,2011,19(1):16-121.
[17] 闵敏,张雪静,马红,等.人脂肪间充质干细胞的原代培养及体外成骨成脂诱导分化[J].江苏大学学报(医学版),2013,23(3):185-190.
[18] 杨柳,朱鸣阳,黑砚,等.人眼眶前脂肪细胞的培养和诱导分化及鉴定[J].医学研究生学报,2011,24(10):57-60.
[1] 刘树林, 温琦, 张永胜, 包斯琴高娃, 王雪, 郭晓宇, 闫素梅.饲粮中非纤维性碳水化合物/中性洗涤纤维对绒山羊羔羊生长性能、屠宰性能及器官指数的影响[J]. 动物营养学报, 2018,30(9): 3543-3550
[2] 马衍旋, 王文杰, 穆淑琴, 席静宁, 李宁, 郑梓, 闫峻, 孙超.共培养体系中猪骨骼肌卫星细胞对肌内前体脂肪细胞脂质沉积的影响[J]. 动物营养学报, 2018,30(4): 1423-1430
[3] 程镇燕, 曲木, 孙颖, 于宏, 孙金辉, 乔秀亭.精氨酸在体内和体外试验中对鲤鱼免疫力的影响[J]. 动物营养学报, 2017,29(9): 3293-3300
[4] 丁志丽, 曹访, 罗娜, 孔有琴, 张易祥, 李景芬, 叶金云.花生四烯酸对日本沼虾肝胰腺细胞脂质代谢基因表达的影响[J]. 动物营养学报, 2017,29(2): 536-546
[5] 李文清, 王加启, 南雪梅, 卜登攀.奶牛乳腺上皮细胞的不同培养方法比较及激素和细胞因子对β-酪蛋白mRNA表达的诱导[J]. 动物营养学报, 2014,26(9): 2607-2614
[6] 刘刚, 任文凯, 熊霞, 李铁军, 钟瑾, 印遇龙.肠道与微生物相互作用体外研究模型进展与应用[J]. 动物营养学报, 2013,25(8): 1677-1682
[7] 成海建, 臧坤, 刘晓牧, 万发春, 谭秀文, 刘桂芬, 宋恩亮.不同培养方法对牛前体脂肪细胞增殖分化的影响[J]. 动物营养学报, 2013,25(4): 864-869
[8] 满意, 张春勇, 陈克嶙, 李美荃, 郭荣富.博落回提取物对脂多糖诱导猪应激细胞应激参数及免疫球蛋白G和超氧化物歧化酶mRNA表达的影响[J]. 动物营养学报, 2012,24(12): 2507-2514
[9] 蔡勇1,2,阿依木古丽3,杨具田3,马忠仁3,卢建雄3,臧荣鑫,吴建平.绵羊前体脂肪细胞的原代培养及分化[J]. 动物营养学报, 2010,22(06): 1768-1774
[10] 孙志洪1,2,张庆丽1,3,贺志雄1,2,韩雪峰1,谭支良1*,张恩平,汤少勋,周传社,王敏.山羊瘤胃上皮细胞和空肠黏膜上皮细胞原代培养技术研究[J]. 动物营养学报, 2010,22(03): 602-610
Copyright 2010 by 动物营养学报