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Primary study of cell phenotype of isolated and cultured canine bone marrow mesenchymal stem cells in vitro
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 20
- DOI:
- 10.3321/j.issn:1000-2790.2005.20.003
- Key Word:
- 间质干细胞;骨髓;组织工程;免疫组织化学;犬
Abstract： 目的: 观察犬骨髓间充质干细胞(BMSCs)经分离、自然扩增及诱导分化后表型变化和生物学特性. 方法: 利用骨髓穿刺、percoll密度梯度法离心分离继而贴壁筛选纯化BMSCs进行接种培养,体外扩增;倒置显微镜下观察其形态学变化,并进行相关抗原检测. 结果: 分离的细胞免疫组化显示SH2, Vimentin, α-SMA和 collagen Ⅰ, Ⅲ均呈阳性表达,其阳性率分别为(95.1±1.7)%, (64.4±2.2)%, (78.7±0.9)%, (78.3±1.4)%和(66.8±0.8)%;CD34,Ⅷ因子antigen 和laminin的表达均为阴性. 在加入bFGF等诱导培养基作用下,可以诱导分化出成纤维细胞,其诱导分化率为(50.2±2.5)%. 诱导后α-SMA表达阳性率下降为(42.3±1.6)%,vimentin表达阳性率上升为(86.2±2.4)%,生长扩增能力强,2 wk内经3代培养即可扩增到(5.6±0.3)×107细胞数量级. 结论: 此实验自然分化扩增来的肌纤维母细胞的表型和生物学特征显示这类间质细胞较应用bFGF诱导、分化扩增来的成纤维细胞有更强活力,且分泌胶原力强,更符合构建组织工程种子细胞的要求.
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