Isolation, Characterization, and Multipotent Differentiation of Mesenchymal Stem Cells Derived from Meniscal Debris.
Isolation, Characterization, and Multipotent Differentiation of Mesenchymal Stem Cells Derived from Meniscal Debris.
复制标题
半月板碎片衍生的间充质干细胞的分离、表征和多能分化。
DOI：
10.1155/2016/5093725
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发表时间：
2016
影响因子：
4.3
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Fu W;Xie X;Li Q;Chen G;Zhang C;Tang X;Li J
本研究旨在培养和表征源自半月板碎片的间充质干细胞。采用酶消化法分离半月板损伤患者半月板碎片中的细胞，体外培养至第三代，通过光镜观察形态和生长情况。还分析了第三代培养物的免疫表型和分化成成骨、脂肪形成和软骨形成谱系的能力。培养4-5天后，细胞呈长纺锤形并粘附在塑料壁上。 10-12天后，观察细胞簇和集落。第三代细胞形态均匀，增殖良好。它们表达 CD44、CD90 和 CD105，但 CD34 和 CD45 呈阴性。通过成骨诱导分化的培养物茜素红染色和碱性磷酸酶活性呈阳性。诱导进行脂肪形成的培养物的油红O染色呈阳性。诱导进行软骨形成的培养物用甲苯胺蓝、阿新蓝和 II 型胶原免疫组织化学染色呈阳性，表明软骨特异性基质。这些结果表明，我们从半月板碎片中培养的细胞是能够沿三个谱系分化的间充质干细胞。这些干细胞可能是半月板再生的宝贵来源。
This study aimed to culture and characterize mesenchymal stem cells derived from meniscal debris. Cells in meniscal debris from patients with meniscal injury were isolated by enzymatic digestion, cultured in vitro to the third passage, and analyzed by light microscopy to observe morphology and growth. Third-passage cultures were also analyzed for immunophenotype and ability to differentiate into osteogenic, adipogenic, and chondrogenic lineages. After 4-5 days in culture, cells showed a long fusiform shape and adhered to the plastic walls. After 10–12 days, cell clusters and colonies were observed. Third-passage cells showed uniform morphology and good proliferation. They expressed CD44, CD90, and CD105 but were negative for CD34 and CD45. Cultures induced to differentiate via osteogenesis became positive for Alizarin Red staining as well as alkaline phosphatase activity. Cultures induced to undergo adipogenesis were positive for Oil Red O staining. Cultures induced to undergo chondrogenesis were positive for staining with Toluidine Blue, Alcian Blue, and type II collagen immunohistochemistry, indicating cartilage-specific matrix. These results indicate that the cells we cultured from meniscal debris are mesenchymal stem cells capable of differentiating along three lineages. These stem cells may be valuable source for meniscal regeneration.
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