阿尔巴斯绒山羊骨髓间充质干细胞体外分离培养及鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2016.02.028

生物技术通报 ›› 2016, Vol. 32 ›› Issue (2): 203-210.doi: 10.13560/j.cnki.biotech.bull.1985.2016.02.028

阿尔巴斯绒山羊骨髓间充质干细胞体外分离培养及鉴定

马苏日古嘎¹, 彭航², 刘佳佳³, 张亦婷⁴, 桂花¹, 刘鹏霞¹

1.内蒙古大学生命科学学院，呼和浩特 010021；2.西安交通大学前沿科学技术研究院骨骼关节疾病与治疗研究中心，西安 710061；3.中国科学院微生物研究所，北京100101；4.中国科学院上海药物研究所药物安全性评价中心，上海 201203

收稿日期:2015-09-06 出版日期:2016-02-24 发布日期:2016-02-25
作者简介:马苏日古嘎，女，硕士研究生，研究方向：干细胞生物学；E-mail：1017955583@qq.com
基金资助:
国家自然科学基金项目(31101698)，内蒙古杰出青年自然科学基金项目(2011JQ03)

Isolation Culture and Identification of Bone Marrow-derived Mesenchymal Stem Cells from Arbas Cashmere Goat

MA Su-ri-gu-ga¹, PENG Hang², LIU Jia-jia³, ZHANG Yi-ting⁴, GUI Hua¹, LIU Peng-xia¹

(1.College of Life Sciences，Inner Mongolia University，Hohhot 010021；2.Bone and Joints Research Center，Frontier Institute of Science and Technology，Xi’an Jiaotong University，Xi’an 710061；3.Institute of Microbiology，Chinese Academy of Sciences，Beijing 100101；4.Center for Drug Safety Evaluation and Research，Shanghai Institute of Materia Medica，Chinese Academy of Sciences，Shanghai 201203)

Received:2015-09-06 Published:2016-02-24 Online:2016-02-25

摘要/Abstract

摘要： 探讨阿尔巴斯绒山羊骨髓间充质干细胞(Arbas cashmere goat bone marrow-derived mesenchymal stem cells，gBM-MSCs)体外分离培养，并对其生物学特性进行系统的鉴定，进一步深入分析体外诱导时多向分化相关基因动态表达情况。结果显示，原代培养的gBM-MSCs呈梭形或纺锤形、放射状排列的细胞集落，能够连续稳定传代至15代以上；免疫荧光及流式细胞术检测显示，gBM-MSCs表达间质系特异性表面标志物，如：CD13、CD29、CD44、CD106、CD90及CD166，不表达造血干细胞表面标志物CD45及CD34；体外诱导实验证实该细胞具有多向分化潜能，能够向脂肪细胞、成骨细胞和软骨细胞分化；荧光定量PCR实验结果显示，gBM-MSCs多向分化相关基因的表达水平随着诱导天数的增加呈上升趋势。实验结果证实阿尔巴斯绒山羊骨髓中分离培养的细胞具备gBM-MSCs的生物学特性。

关键词: 阿尔巴斯绒山羊, 间充质干细胞, 培养, 分化, 生物学特性

Abstract: Here we isolated and cultured Arbas cashmere goat bone marrow-derived mesenchymal stem cells(gBM-MSCs)in vitro, identified the biological characteristics of gBM-MSCs, and analyzed the dynamic expressions of differentiation genes while induced in vitro.The primary cultured gBM-MSCs were spindle cell-based, showing radial colony arrangement, and stably and continuously passed down for over 15 generations.The detections by immunofluorescence and flow cytometry showed that gBM-MSCs were positive for mesenchymal specific markers, such as CD13, CD29, CD44, CD106, CD90 and CD166, but negative for hematopoietic stem cell surface markers CD45 and CD34.The induction in vitro confirmed that gBM-MSCs owned the multiple differentiation potential, and differentiated into adipocytes, osteoblasts and chondrocyte.Real-time PCR further indicated that the expressions of differentiation genes increased throughout the culture period.Our results confirmed that the isolated and cultured cells from the bone marrow of Arbas cashmere goat possessed the biological characteristics of gBM-MSCs.Our findings promoted the basic research of MSCs in livestock and laid a theoretical foundation for the further application of gBM-MSCs in transgenic breeding and tissue engineering.

Key words: Arbas cashmere goat, mesenchymal stem cells, culture, differentiate, biological characteristics

马苏日古嘎, 彭航, 刘佳佳, 张亦婷, 桂花, 刘鹏霞. 阿尔巴斯绒山羊骨髓间充质干细胞体外分离培养及鉴定[J]. 生物技术通报, 2016, 32(2): 203-210.

MA Su-ri-gu-ga, PENG Hang, LIU Jia-jia, ZHANG Yi-ting, GUI Hua, LIU Peng-xia. Isolation Culture and Identification of Bone Marrow-derived Mesenchymal Stem Cells from Arbas Cashmere Goat[J]. Biotechnology Bulletin, 2016, 32(2): 203-210.

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阿尔巴斯绒山羊骨髓间充质干细胞体外分离培养及鉴定

Isolation Culture and Identification of Bone Marrow-derived Mesenchymal Stem Cells from Arbas Cashmere Goat

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