Li Linfeng, Bai Xiujuan, Gong Xuelian, Liu Hongkun, Chen Lina, Guan Weijun, Ma Yuehui
Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, China.
J Genet Genomics. 2009 Mar;36(3):133-40. doi: 10.1016/S1673-8527(08)60100-9.
The bone marrow mesenchymal stem cells (MSCs) are multipotent stem cells which can differentiate into mesenchymal cells in vitro. In this study, MSCs in duck were isolated from bone marrow by density gradient centrifuge separation, purified and expanded in the medium. The primary MSCs were expanded for passages. The different-passage MSCs were induced to differentiate into osteoblasts and neuron-like cells. Karyotype analysis indicated that MSCs kept diploid condition and the hereditary feature was stable. The different-passage MSCs expressed CD44, ICAM- and SSEA-4, but not CD34, CD45 and SSEA-when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 2, 6 and 8 (P > 0.05), but a significant difference existed among those of passages 2, 6, 8 and 11 (P < 0.05). After the osteogenic inducement was added, the induced different-passage MSCs expressed high-level alkaline phosphatase (ALP), and are positive for tetracycline staining, Alizarin Red staining and Von Kossa staining. After the neural inducement was added, about 70% cells exhibited typical neuron-like phenotype, the induced different-passage MSCs expressed Nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 3, 4 and 6 (P>0.05), but a significant difference existed among those of passages 3, 4, 6 and 8 (P<0.05). These results suggest that MSCs in duck were capable of differentiating into osteoblasts and neuron-like cells in vitro.
骨髓间充质干细胞（MSCs）是多能干细胞，可在体外分化为间充质细胞。在本研究中，通过密度梯度离心分离从鸭骨髓中分离出MSCs，在培养基中进行纯化和扩增。原代MSCs进行传代培养。对不同传代的MSCs诱导分化为成骨细胞和神经元样细胞。核型分析表明MSCs保持二倍体状态，遗传特征稳定。免疫荧光染色检测显示，不同传代的MSCs表达CD44、ICAM-和SSEA-4，但不表达CD34、CD45和SSEA-。第2、6和8代的阳性率之间无显著差异（P>0.05），但第2、6、8和11代之间存在显著差异（P<0.05）。添加成骨诱导剂后，诱导的不同传代MSCs表达高水平碱性磷酸酶（ALP），对四环素染色、茜素红染色和冯科萨染色呈阳性。添加神经诱导剂后，约70%的细胞呈现典型的神经元样表型，免疫荧光染色检测显示，诱导的不同传代MSCs表达巢蛋白、神经元特异性烯醇化酶（NSE）和胶质纤维酸性蛋白（GFAP）。第3、4和6代的阳性率之间无显著差异（P>0.05），但第3、4、6和8代之间存在显著差异（P<0.05）。这些结果表明鸭MSCs能够在体外分化为成骨细胞和神经元样细胞。
