[1]王家敏,沈武玲,平 玲,等.婴儿皮肤组织成纤维细胞系的建立及生物学特性[J].南京师大学报(自然科学版),2014,37(03):100.
　Wang Jiamin,Sheng Wuling,Ping Ling,et al.Establishment and Characteristics of Baby Skin Explant Fibroblast[J].Journal of Nanjing Normal University(Natural Science Edition),2014,37(03):100.
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婴儿皮肤组织成纤维细胞系的建立及生物学特性()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第37卷
期数:
2014年03期
页码:
100
栏目:
生命科学
出版日期:
2014-09-30

文章信息/Info

Title:
Establishment and Characteristics of Baby Skin Explant Fibroblast
作者:
王家敏1; 沈武玲2; 平 玲2; 徐水林1; 赵 彬2; 马忠仁1; 乔自林1
(1.甘肃省动物细胞工程技术研究中心,兰州 730030)(2.西北民族大学生命科学与工程学院,兰州 730030)
Author(s):
Wang Jiamin1; Sheng Wuling2; Ping Ling2; Xu Shuilin1; Zhao Bin2; Ma Zhongren1; Qiao Zilin1
(1.Gansu Engineering Research Center for Animal Cell,Lanzhou 730030,China)(2.College of Life Science and Engineering,Northwest University for Nationalities,Lanzhou 730030,China)
关键词:
婴儿皮肤; 成纤维细胞系; 生物学特性
Keywords:
baby skin; fibroblast cell line; characteristics
分类号:
Q813.1+1
文献标志码:
A
摘要:
采集婴儿皮肤组织,用组织块法制备原代细胞,通过差速消化和差速贴壁法纯化细胞,扩大培养至第3代液氮冻存,复苏细胞后进行活力、形态、生长曲线、微生物污染、核型、乳酸脱氢同工酶谱及荧光蛋白质粒转染表达等特性研究.结果显示,婴儿皮肤组织细胞呈成纤维型,生长状态良好; 生长曲线呈“S”型,最大增值浓度为1.64×105/mL,倍增时间为63.12 h; 细菌、真菌、支原体检测均为阴性; 染色体2n=46,二倍体为86%,占主体; 乳酸脱氢酶同工酶电泳图谱显示有5条谱带; 外源质粒在该细胞中能进行复制和表达.表明已成功建立了婴儿皮肤组织成纤维细胞系,这为复合人工皮肤的组织工程学研究奠定了前期工作基础,也为皮肤相关疾病的研究提供了实验条件,因而具有重要的理论和实践意义.
Abstract:
The baby skin explants were cultured with tissue pieces method.The primary cells were purified and preserved with liquid nitrogen after being passaged to the thrid generation.Observations on morphology,dynamic growth,microbial contamination,analysis of karyotype,isoenzyme of lactate dehydrogenase and fluorescin plasmid transfection and expression were carried out.The results showed that the cell line was fibroblast and growed well.The maximum proliferation concentration was 1.64×105/mL and poulation doubling time of cells(PDT)was 63.12 h.The tests for bacteria,fungi,virus and mycoplasma were all negative.The chromosomes were 46 and diploid cells were dominant of 86%.The banding patterns of the isoenzyme of lactate dehydrogenase had significant difference and there were five bands in all.The exogenous plasmid could copy and express in the cell.Those showed that the cell line was established successfully,which lays the foundation of the work bases for composite artificial skin tissue engineering and provides the experimental conditions for skin diseases.So this has the important theoretical and practical significance.

参考文献/References:

[1] 吴国选,伍津津,朱堂友,等.人皮肤成纤维细胞库的构建[J].实用医药杂志,2005,22(4):324-326.
[2]侯兰新,陈轶霞.动物细胞培养技术教程[M].兰州:甘肃科学技术出版社,2009:58-59,124-126.
[3]沈翠英,肖忠明.原代培养的二倍体上皮细胞和成纤维细胞的纯化[J].细胞生物学杂志,1989,11(4):173-174.
[4]刘永清,孙浩.鸡胚成纤维细胞原代培养与纯化的初探[J].猪与禽,2008,28(1):72-74.
[5]程宝鸾.动物细胞培养技术[M].广州:中山大学出版社,2006:28-29.
[6]李玲,李雪峰.细胞生物学实验[M].长沙:湖南科学技术出版社,2003:17-19.
[7]严泉剑,郭金龙,刘恩靖,等.绘制细胞生长曲线及细胞群体倍增时间的简化计算[J].前卫医药杂志,2000,17(4):228-229.
[8]国家药典委员会.中华人民共和国药典(2005年版)[M].北京:化学工业出版社,2005:14-15,75-77.
[9]雷初朝,韩增胜,陈宏,等.关于马的染色体核型分析[J].西北农林科技大学学报,2001,29(4):5-8.
[10]鄂征.组织培养和分子细胞学技术[M].北京:北京出版社,2001:164-169.
[11]Yunis J J.Human Chromosome Methodology[M].New York:Academic Press,1974:59-61.
[12]Franklin E,William F,Blazak.Horse,ass,and mule chromosomes[J].Journal of Heredity,1976,67:361-367.
[13]良宋平.生物化学与分子生物学实验教程[M].北京:高等教育出版社,2003:35-41.
[14]管卫,齐静,沈永明,等.1例儿童血清乳酸脱氢酶同工酶异常区带的分析[J].临床检验杂志,2010,28(1):75-76.
[15]周向梅,马月辉,关伟军,等.北京油鸡胚胎成纤维细胞系建立与生物学特性研究[J].畜牧兽医学报,2005,36(3):209-215.
[16]Drexel H G,Dirks W G,Macl E R,et al.Sensitivity hematopoietic cell lines:cross-contaminations and misinterpretations[J].Keukemia,1999,13:602-606.
[17]何申.ATCC美国所属培养的细胞库[M].北京:中国学术期刊电子杂志社,2006:52-56.

备注/Memo

备注/Memo:
收稿日期:2012-08-28.
基金项目:西北民族大学中青年科研基金.
通讯联系人:乔自林,高级实验师,研究方向:细胞培养技术和细胞生物反应器.E-mail:670267497@qq.com
更新日期/Last Update: 2014-09-30