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| 三种不同型别小鼠骨髓来源间质干细胞的生物学特性分析 | |
| 引用本文： | 惠大阳,郭振宇,卢志杰,余伟华,王涛,雷俊霞.三种不同型别小鼠骨髓来源间质干细胞的生物学特性分析[J].中山大学学报(医学科学版),2011,32(2). |
| 作者姓名： | 惠大阳 郭振宇 卢志杰 余伟华 王涛 雷俊霞 |
| 作者单位： | 1. 中山大学,中山大学干细胞与组织工程中心,广东,广州,510080;中山大学,中山医学院病理生理教研室,广东,广州,510080 2. 附属第五医院泌尿外科,广东,珠海,519000 3. 广东医学院附属第一医院放疗科,广东,广州,510080 4. 中山大学,中山大学干细胞与组织工程中心,广东,广州,510080 |
| 摘 要： | 【目的】间质干细胞(MSC)是一种间质来源的多潜能基质细胞，目前人和多种动物来源的MSC均已被成功分离鉴定。然而由于小鼠骨髓MSC的分离相对人和其他物种更为困难，关于小鼠骨髓MSC的克隆分析结果也相对有限且结果不尽一致。为此,本研究拟进一步探讨小鼠MSC的体外分离方法，并对MSC克隆形成单位进行生物学特性分析。【方法】 取C57BL/6小鼠，冲洗股骨骨髓腔获得骨髓单个核细胞悬液，低密度接种培养,并通过有限稀释克隆挑选获得三种形态、生长特点不同的克隆形成单位(CFU)。采用流式分析技术对三种类型的CFU进行表型分析，并用油红O和茜素红分别进行成脂和成骨分化诱导鉴定。【结果】 低密度培养并结合克隆化培养分离技术成功获得C57BL/6小鼠骨髓间充质干细胞,并观察到偏圆形(MSC1)、长梭形(MSC2)以及纺锤形、多边形(MSC3)三种贴壁形态细胞类型；免疫表型分析显示，三种细胞均强表达Sca-1，不表达CD11b、CD45，部分表达CD90.2；体外诱导分化实验证实，MSC1仅具有向脂肪细胞分化的潜能，MSC2仅具有向成骨细胞分化的潜能;MSC3则具有成骨、成脂双向分化能力。【结论】 低密度培养并结合克隆化培养分离技术可成功分离小鼠骨髓间充质干细胞;小鼠MSCs是一种高度异质性的细胞群，其中可能含有多能MSC或单一分化方向的前体细胞等处于不同分化阶段的细胞类型。 |
| 关 键 词： | 骨髓 间充质干细胞 小鼠 生物学特性 |
| 收稿时间： | 2011-01-07; |
Biological Characteristics Analysis of Three Types of Murine Bone Marrow Mesenchymal Stem Cells |
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| HUI Da-yang,GUO Zhen-yu,LU Zhi-jie,YU Wei-hua,WANG Tao,LEI Jun-xia.Biological Characteristics Analysis of Three Types of Murine Bone Marrow Mesenchymal Stem Cells[J].Journal of Sun Yatsen University(Medical Sciences),2011,32(2). | |
| Authors: | HUI Da-yang GUO Zhen-yu LU Zhi-jie YU Wei-hua WANG Tao LEI Jun-xia |
| Affiliation: | HUI Da-yang1,2,GUO Zhen-yu3,LU Zhi-jie4,YU Wei-hua1,WANG Tao1,LEI Jun-xia1,2*(1.Center for Stem Cell Biology and Tissue Engineering,Sun Yat-sen University,Guangzhou 510080,China,2.Department of Pathophysiology,Zhongshan Medical College,3.Department of Urinary Surgery,The Fifth Affiliated Hospital,Zhuhai 519000,4.Department of Radiotherapy,The First Affiliated Hospital of Guangdong University of Pharmacy,China) |
| Abstract: | 【Objective】 Mesenchymal stem cells (MSC) are multipotential stromal cells. Now the MSC of human and many animals have been isolated and identified successfully. However, the isolation of purified murine MSC from bone marrow is far more difficult than that in human and other species, so the clonal analysis results of murine bone marrow MSC are limited and even inconsistent. Therefore, isolation of pure murine MSC in vitro and identification of the biological characteristics of colony-forming units(CFU)were further studied. 【Methods】 Mononuclear cells flushed from thigh bone marrow cavity of C57BL/6 mouse were plated at low density. Three types of CFU with different morphology and proliferation characteristics were acquired by limiting dilution and cloning selection. The immunophenotype of these three types of CFU were analyzed by flow cytometry and the osteogenic differentiation and adipocytic differentiation potential were identified by Alizarin red staining and Oil Red O staining. 【Results】 Thus, by low density culture system combined with cloning isolation technique, we totally obtained three distinct clonal MSC populations from C57BL/6 mouse bone marrow. These cell lines exhibited different morphology: MSC1 had a near round morphology, while MSC2 presented a typical fibroblast-like morphology and MSC3 had fusiform and polygonal morphology. These three cell populations were all strong positive for stem cell antigen-1 (Sca-1), negative for CD11b and CD45 and partially positive for CD90.2. Differentiation potential assay showed that MCS1 could only be induced to adipocyte and MSC2 only to osteoblast, while MSC3 could differentiate to adipocyte and osteoblast.【Conclusions】 Low density culture system combined with cloning isolation technique could isolate mouse mesenchymal stem cells successfully. Mouse MSC are highly heterogeneous cell populations which may contain multi-potential MSC or uni-potential progenitors at different developmental stages. |
| Keywords: | bone marrow mesenchymal stem cells murine biological characteristics |
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