北京油鸡胚胎肝脏来源间充质干细胞的分离培养及生物学特性

doi:10.3724/SP.J.1005.2013.00365

遗传 ›› 2013, Vol. 35 ›› Issue (3): 365-372.doi: 10.3724/SP.J.1005.2013.00365

北京油鸡胚胎肝脏来源间充质干细胞的分离培养及生物学特性

牧仁^1,2, 边艳超^1,2, 浦亚斌², 李向臣², 王凤龙¹, 关伟军²

1. 内蒙古农业大学兽医学院, 呼和浩特 010018 2. 中国农业科学院北京畜牧兽医研究所, 北京 100193

收稿日期:2012-08-31 修回日期:2012-10-10 出版日期:2013-03-20 发布日期:2013-03-25
通讯作者: 王凤龙 E-mail:wangfengloo@sohu.com
基金资助:
转基因生物新品种培育科技重大专项(编号：2011ZX08009-003-006, 2011ZX08012-002-06), 基本科研业务项目(编号：2012z1072)和国家家养动物种质资源平台项目(2012年)资助

Isolation and biological characterization of mesenchymal stem cells from Beijing fatty chicken fetal liver

MU Ren^1,2, BIAN Yan-Chao^1,2, PU Ya-Bin², LI Xiang-Chen², WANG Feng-Long¹, GUAN Wei-Jun²

1. College of Animal Veterinary, Inner Mongolia Agricultural University, Hohhot 010018, China 2. Institute of Beijing Animal Science and Veterinary, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2012-08-31 Revised:2012-10-10 Online:2013-03-20 Published:2013-03-25

摘要/Abstract

摘要： 鸡胎肝间充质干细胞是一种多能干细胞, 可以在体外分化成多种终末分化细胞; 目前大多数肝脏间充质干细胞研究都集中在大鼠和小鼠, 其他动物研究相对较少。文章从7日龄北京油鸡胚胎肝脏中分离间充质干细胞, 原代培养并传代至15代。免疫荧光法检测间充质干细胞表面标志物CD29、CD44呈阳性表达, 同时检测造血祖细胞/肝卵圆细胞的表面标志物CD34、CK19呈阴性表达。RT-PCR检测CD29、CD44、CD71、CD73呈阳性表达。细胞生长曲线呈“S”型。肝脏间充质干细胞通过不同诱导液被成功诱导分化成神经细胞、成骨细胞。结果表明, 从鸡胎肝中分离获得的间充质干细胞具有和小鼠的间充质干细胞相似的生物学特性及分化潜能, 其具有多向分化的潜能为今后临床广泛应用提供了可能行。

关键词: 北京油鸡, 间充质干细胞, 增殖能力, 诱导分化

Abstract: Mesenchymal stem cells (MSCS) from chicken fetal liver are multipotent stem cells that can differentiate in vitro into various terminally differentiated cells. The majority of studies have focused on rats and mice now. Reports from other animals are less and analyses on domestic animals are few. In this study, chicken liver-derived MSCs were isolated from 7-day-old embryo of Beijing fatty chickens. Primary liver-derived MSCs were subcultured to passage 15. The surface markers of liver-derived MSCs, CD29, and CD44 were detected by immunofluorescence and the surface markers CD34 and CK19 of hematopoietic progenitor cells/hepatic oval cells were not detected. RT-PCR analysis detected positive expression of CD29, CD44, CD71, and CD73. The growth curves were typically sigmoidal. Liver-derived MSCs of different passages were successfully induced and differentiated into neuronal and osteoblast cells. The results suggest that the MSCs isolated from chicken fetal liver possess similar biological characteristics with those derived from mice, and their multilineage dif-ferentiation provides many potential applications.

Key words: Beijing fatty chicken, mesenchymal stem cells, proliferation, differentiation potential

牧仁，边艳超，浦亚彬，李向臣，王凤龙，关伟军. 北京油鸡胚胎肝脏来源间充质干细胞的分离培养及生物学特性[J]. 遗传, 2013, 35(3): 365-372.

MU Ren BIAN Yan-Chao PU Ya-Bin LI Xiang-Chen WANG Feng-Long GUAN Wei-Jun. Isolation and biological characterization of mesenchymal stem cells from Beijing fatty chicken fetal liver[J]. HEREDITAS, 2013, 35(3): 365-372.

参考文献

[1] 罗时和. 北京油鸡. 农业知识, 2008, (8): 46.
[2] 马月辉, 徐桂芳, 王端云, 刘海良. 中国畜禽遗传资源信息动态研究. 中国农业科学, 2002, 35(5): 552-555.
[3] 陈幼春, 马月辉, 何晓红, 浦亚斌. 地方、培育、引入品种资源的保存与发展的研究进展. 中国畜牧兽医, 2008, 35(1): 5-11.
[4] 刘刚. 国家级畜禽遗传资源保护名录. 四川农业科技, 2007, (5): 42.
[5] Friedenstein AJ, Petrakova KV, Kurolesova AI, Frolova GP. Heterotopic of bone marrow. Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation, 1968, 6(2): 230-247.
[6] 王恒湘, 郭子宽. 间充质干细胞在组织再生应用中的诸多问题. 组织工程与重建外科杂志, 2008, 4(5): 241-245.
[7] Jenkins N. Animal cell biotechnology: Methods and protocols. New Jersey: Humana Press, 1999: 132-138.
[8] Tan J, Liu T, Hou L, Meng WT, Wang YC, Zhi W, Deng L. Maintenance and expansion of hematopoietic stem/progenitor cells in biomimetic osteoblast niche. Cytotechnology, 2010, 62(5): 439-448.
[9] Honmou O, Houkin K, Matsunaga T, Niitsu Y, Ishiai S, Onodera R, Waxman SG, Kocsis JD. Intravenous admini-stration of auto serum-expanded autologous mesenchymal stem cells in stroke. Brain, 2011, 134(6): 1790-1807.
[10] 周向梅, 马月辉, 关伟军, 文杰, 李晗. 北京油鸡胚胎成纤维细胞系建立与生物学特性研究. 畜牧兽医学报, 2005, 36(3): 209-215.
[11] Erices A, Conget P, Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood. Brit J Haematol, 2000, 109(1): 235-242.
[12] Seong JM, Kim BC, Park JH, Kwon K II, Mantalaris A, Hwang YS. Stem cells in bone tissue engineering. Biomed Mater, 2010, 5(6): 062001.
[13] Girdlestone J, Limbani VA, Cutler AJ, Navarrete CV. Efficient expansion of mesenchymal stromal cells from um-bilical cord under low serum conditions. Cytotherapy, 2009, 11(6): 738-748.
[14] 刘伟, 刘萌, 祝劲松, 郝红伟, 董宁征, 周海滨. 人骨髓间充质干细胞的体外培养、鉴定及成骨分化. 中国组织工程研究, 2102, 16(14): 2515-2519.
[15] Birmingham E, Niebur GL, McHugh PE, Shaw G, Barry FP, McNamara LM. Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche. Eur Cell Mater, 2012, 12(23): 13-27.
[16] 余卫, 张洹, 何冬梅. 体外诱导小鼠胎肝间充质干细胞向胰岛B样细胞分化的研究. 第二军医大学学报, 2004, 25(8): 827-830.
[17] Haasters F, Prall WC, Anz D, Bourquin C, Pautke C, Endres S, Mutschler W, Docheva D, Schieker M. Mor-phological and immunocytochemical characteristics indi-cate the yield of early progenitors and represent a quality control for human mesenchymal stem cell culturing. J Anat, 2009, 214(5): 759-767.
[18] Colter DC, Class R, DiGirolamo CM, Prockop DJ. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci USA, 2000, 97(7): 3213-3218.
[19] Morikawa S, Mabuchi Y, Kubota Y, Nagai Y, Niibe K, Hiratsu E, Suzuki S, Miyauchi-Hara C, Nagoshi N, Sunabori T, Shimmura S, Miyawaki A, Nakagawa T, Suda T, Okano H, Matsuzaki Y. Prospective identification, iso-lation, and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow. J Exp Med, 2009, 206(11): 2483-2496.
[20] 郭子宽. 间充质干细胞及其临床应用中的几个问题. 中国组织工程研究, 2012, 16(1): 1-10.
[21] Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 2006, 126(4): 663-676.
[22] Jiang RH, Han ZB, Zhou GS, Qu XD, Li X, Wang X, Shao YK, Yang SM, Han ZC. Transplantation of placenta-derived mesenchymal stem cells in type 2 diabetes: a pilot study. Front Med, 2011, 5(1): 94-100.
[23] Nakajima H, Uchida K, Guerrero AR, Watanabe S, Sugita D, Takeura N, Yoshida A, Long G, Wright KT, Johnson WE, Baba H. Transplantation of mesenchymal stem cells promotes an alternative pathway of macrophage activation and functional recovery after spinal cord injury. J Neurotrauma, 2012, 29(8): 1614-1625.

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北京油鸡胚胎肝脏来源间充质干细胞的分离培养及生物学特性

Isolation and biological characterization of mesenchymal stem cells from Beijing fatty chicken fetal liver

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