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鸡骨髓间充质干细胞的分离与鉴定

Isolation and characterization of mesenchymal stem cells from chicken bone marrow.

作者信息

Bai Chunyu, Hou Lingling, Ma Yuehui, Chen Lina, Zhang Minghai, Guan Weijun

机构信息

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, People's Republic of China.

出版信息

Cell Tissue Bank. 2013 Sep;14(3):437-51. doi: 10.1007/s10561-012-9347-8. Epub 2012 Nov 16.

Abstract

The bone marrow mesenchymal stem cells (BMSCs) are multipotent stem cells, which can differentiate in vitro into many cell types. However, the vast majority of experimental materials were obtained from human, mouse, rabbit and other mammals, but rarely in poultry. So, in this study, Thirty- to sixty-day old chicken was chosen as experimental animal, to isolate and characterize BMSCs from them. To investigate the biological characteristics of chicken BMSCs, immunofluorescence and RT-PCR were used to detect the characteristic surface markers of BMSCs. Growth curves were drawn in accordance with cell numbers. To assess the differentiation capacity of the BMSCs, cells were induced to differentiate into osteoblasts, adipocytes, and endothelial cells. The surface markers of BMSCs, CD29, CD44, CD31, CD34, CD71 and CD73, were detected by immunofluorescence and RT-PCR assays. The growth curves of different passages were all typically sigmoidal. Karyotype analysis showed that these in vitro cultured cells were genetically stable. In addition, BMSCs were successfully induced to differentiate into osteoblasts, adipocytes, and endothelial cells. The results suggest that the BMSCs isolated from chicken possess similar biological characteristics with those separated from other species, and their multi-lineage differentiation potentiality herald a probable application for cellular transplant therapy in tissue engineering.

摘要

骨髓间充质干细胞(BMSCs)是多能干细胞,能够在体外分化为多种细胞类型。然而,绝大多数实验材料取自人、小鼠、兔子和其他哺乳动物,在家禽中却很少见。因此,在本研究中,选用30至60日龄的鸡作为实验动物,从中分离并鉴定BMSCs。为了研究鸡BMSCs的生物学特性,采用免疫荧光和RT-PCR检测BMSCs的特征性表面标志物。根据细胞数量绘制生长曲线。为了评估BMSCs的分化能力,将细胞诱导分化为成骨细胞、脂肪细胞和内皮细胞。通过免疫荧光和RT-PCR检测BMSCs的表面标志物CD29、CD44、CD31、CD34、CD71和CD73。不同传代的生长曲线均呈典型的S形。核型分析表明,这些体外培养的细胞基因稳定。此外,BMSCs成功诱导分化为成骨细胞、脂肪细胞和内皮细胞。结果表明,从鸡分离的BMSCs与从其他物种分离的BMSCs具有相似的生物学特性,其多谱系分化潜能预示着在组织工程细胞移植治疗中可能具有应用价值。

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