2009---广西巴马香猪耳皮成纤维细胞的分离与培养

第3期(责任编辑王珞)

参考文献:

[1]

SHEN M M ,HODGKIN J.Mab-3,a gene required for sex specific yolk protein expression and a male specific lineage in C.elegans [J ].Cell ,1988,54:1019-1031.

[2]BURTUS K C ,BAKER B S.Drosophila double sex gene control somatic sexual differentiation by producing alteratively spliced mRNAs encoding related sex specific polypeptides [J ].Cell ,1989,56:997-1010.

[3]RAYMOND C S ,SHAMU C E ,SHEN M M ,et a1.Evi-dence for evolutionary conservation of sex determining genes [J ].Nature ,1998,391:691-695.

[4]GRANDI A D ,CALVARI V ,BERTINI V ,et a1.The ex-pression pattern of a monse doublesex ·related gene is con-sistent with a role in gonadal differentiation [J ].Mech Dev ,

2000,90:323-326.[5]

REN L L ,CHENG H H ,GUO Y Q ,et a1.Evolutionary conservation of Dmrt gene family in amphibians reptiles and birds [J ].Chin Sci Bull ,2001,46(23):1992-1996.

[6]姚凯,赵洋,张涛,等.哺乳动物DMRT1基因调控区分

析[J ].遗传,2006,28(7):825-830.

[7]

RAYMOND C S ,PARKER E D ,KETTLEWELL J R ,et a1.A region of human chromosome 9p required for testis de-velopment contains two genes related to known sexual regula-tors [J ].Hum Molec Genet ,1999,8:989-996.

[8]

PASK A J ,BEHRINGER R R ,RENFREE M B.Expression of DMRT1in the mammalian ovary and testis-from marsupi-als to mice [J ].Genome Res ,2003,101:229-236.

第48卷第3期2009

3

月湖北农业科学

H ubei A gricultural S ciences

Vol.48No.3Mar .,2009

收稿日期:2009-01-09

基金项目:国家高技术研究发展计划(863)项目(2007AA100505)

作者简介:杨素芳(1972-),女,广西兴安人,副教授,博士,(电话)137********(电子信箱)ysfang3511@163.com ;通讯作者,石德顺。

广西巴马香猪耳皮成纤维细胞的分离与培养

杨素芳,汪彩珠,陈凌声,覃兆鲜,黄雅琼,石德顺

(广西大学动物繁殖研究所,南宁

530005)

摘要:试验主要对广西巴马香猪耳皮成纤维细胞的分离和培养进行了研究。利用组织块法能够成功分离培养广西巴马香猪耳皮成纤维细胞,其大小范围在1~3mm 3,去除表皮有利于组织块贴壁和细胞的游出;而采用0.25%胰蛋白酶消化液4℃冷处理过夜,不仅有利于去表皮操作,减少取材处理时间,而且操作时间自由度大。在酶消化法中采用胰蛋白酶消化30min ,胶原酶继续消化30~40min ,能够成功分离培养广西巴马香猪耳皮成纤维细胞。广西巴马香猪皮肤成纤维细胞要经历滞留期、对数期、平台期,分别为

0~2d 、2~8d 、8~12d 。

关键词:广西巴马香猪;成纤维细胞;体外培养;分离中图分类号:S828.9+9(674BM );Q954.6-33+2

文献标识码:A

文章编号:0439-8114(2009)03-0547-06

Dissociation and Culture of Ear Skin Fibroblasts in Guangxi Bama Miniature Pigs

YANG Su-fang ,WANG Cai-zhu ,CHEN Ling-sheng ,QIN Zhao-xian ,HUANG Ya-qiong ,SHI De-shun

(Animal Reproduction Institute ,Guangxi University ,Nanning 530005,China )

Abstract :Dissociation and culture of ear skin fibroblasts in Guangxi Bama m iniature pigs was studied.Tissue piece culture could dissociate ear skin fibroblast in Guangxi Bama m iniature pigs successfully.Tissue piece with removaling epidermis and the size of 1~3mm 3were contributed to cell swimming out and cell adherence.Tissue piece undergoing 0.25%trypsin cold treatment overnight was helpful to remove its epidermis ,decrease the time of treating material and to operate freely.The fi-broblasts could be disassociated and cultivated successfully by adopting trypsin digestion for 30min firstly and collagenase digestion for 30~40min continuously.The detention period ,log phase and platform time of the ear skin fibroblast in Guangxi Bama m iniature pigs was 0~2d ays ,2~8d ays ,8~12d ays respectively.Key words :

Guangxi Bama m iniature pig ;fibroblast ;in vitro culture ;disassociation

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!

相关推荐
相关主题
热门推荐