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C57BL/6J×129/J杂交小鼠ES细胞系的建株
  • ISSN号:1000-4718
  • 期刊名称:《中国病理生理杂志》
  • 时间:0
  • 分类:R363[医药卫生—病理学;医药卫生—基础医学]
  • 作者机构:[1]中山大学干细胞与组织工程中心,广东广州510089
  • 相关基金:This work was supported by the Major State Basic Research Development Program of China (973 Program, No. 2001CB509904) ; the Key Scientific and Technological Projects of Gnangdong Province (No. 2003A3020103) ; National Natural Science Foundation of China (No 30100188, 30571891) ; We also thank Bruce T Lahn from University of Chieago for his antibodies.
中文摘要:

目的:建立C57BL/6J×129/J杂交小鼠ES细胞系。方法:收集3.5d.p.c.的囊胚,培养在预先铺有小鼠成纤维细胞(MEFs)的高糖DMEM培养液中。3—4d后,挑出内细胞团(ICM),消化后重新种到新鲜的有MEPS培养液中。等到有典型的ES样集落长出,即传代以得到永久ES细胞系。通过分析碱性磷酸酶活性,SSEA-1,Oct-4的表达和形成畸胎瘤的能力来鉴定ES细胞的多向分化能力。结果:获得的两个C57BL/6J×129/J杂交小鼠ES细胞系绝大多数细胞具有正常的核型(40,XY),碱性磷酸酶染色阳性,SSEA—l,Oct-4表达阳性,ES细胞注入SCID鼠后可获得来自3个胚层的组织。结论:建立了两株具有长期自我更新能力和多向分化潜能的C57BL/6J×129/J杂交小鼠ES细胞系。

英文摘要:

AIM: To establish hybrid mouse embryonic stem (KS) cell line from blastocysts of the (C57BL/6J × 129/ J) F1 mouse. METHODS:3.5 days post- coitus (d. p.c. ) blastocysts were cultured on mouse embryonic fibroblasts (MEFs) in the medium, after 3 - 4 days, Inner cell mass were picked up and disaggregated, then reseeded. After the KS - like colonies appeared, passaged them to give permanent KS cell lines. The pluripotent properties of KS cells obtained were analyzed by alkaline phosphatase (AKP) activity, expression of SSEA- 1 and Oct- 4, and their capacity to form teratoma. RESULTS: Two hybrid KS cell lines, SC1001, SC1002 were obtained from blastocysts of the (C57BL/6J × 129/J) F1 genotype. Most of these KS cells had a normal karyotype and an XY sex chromosome composition. The pluripotent properties of the cell lines were analyzed on the basis of their alkaline phosphatase activity, expression of SSEA - 1 and Oct - 4, and their capacity to form teratoma in severe combined immunodeficiency (SCID) mice. CONCLUSION: Two hybrid mouse KS cell lines having pluripotent properties and capacity for long - term self renewal were generated from blastocysts of the (C57BL/6J × 129/J) F1 genotype.

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期刊信息
  • 《中国病理生理杂志》
  • 中国科技核心期刊
  • 主管单位:中国科学技术协会
  • 主办单位:中国病理生理学会
  • 主编:陆大祥
  • 地址:广东省广州市黄埔大道西601号
  • 邮编:510632
  • 邮箱:obsbjbb@jnu.edu.cn
  • 电话:020-85220269
  • 国际标准刊号:ISSN:1000-4718
  • 国内统一刊号:ISSN:44-1187/R
  • 邮发代号:46-98
  • 获奖情况:
  • 1997-2000年连续获得中国科协优秀基础性和高科技...,1992、1996、2000、2004、2008年,连续五次入选中...,2008-2010年,连续三年荣获“百种中国杰出学术期...,2010年获广东省期刊最高奖——品牌期刊奖
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  • 被引量:37010