| 李林蔚 李国庆 胡劲松 谢红艳 罗莎莎 王燕 易岚.小鼠唾液腺干细胞体外分离培养方法研究[J].,2015,15(32):6230-6233 |
| 小鼠唾液腺干细胞体外分离培养方法研究 |
| A Method for Isolation of Mouse Salivary Gland StemCells |
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| DOI： |
| 中文关键词: 小鼠唾液腺干细胞 细胞分离培养 悬浮唾液球 层连蛋白 |
| 英文关键词: Mouse salivary gland stemcells Cell culture techniques Salispheres Laminin |
| 基金项目:国家自然科学基金项目（81400117）；中国博士后基金项目（2014M562115）；湖南省自然科学基金项目（2015JJ4042）；
南华大学归国人员科研启动基金项目（2014XQD46） |
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| 中文摘要: |
| 目的：建立小鼠唾液腺干细胞体外分离培养方法。方法：从7~8 周龄的C57BL/6J 小鼠唾液腺分离唾液腺干细胞，采用悬浮唾
液球法培养唾液腺干细胞，应用倒置显微镜动态观察细胞形态；细胞计数观察细胞生长状况；免疫荧光检测LN（层连蛋白）的表
达。结果：倒置显微镜下，接种2 d后细胞（salispheres）小聚集体明显增加；接种4 d后唾液腺球直径逐渐增大，形成直径约100
滋m的神经球，球周围无明显刺突，存在明显细胞聚集。接种7 d后唾液腺球周围存在较多微刺，10 d后球继续增长，形状明显不同
于4 d，表明处于活跃的增殖状态，唾液腺干细胞已经开始分化。生长曲线结果显示随着培养天数的增加，唾液腺球的数目呈明显
递增趋势，同时，唾液腺球的直径也明显增大。免疫荧光结果显示，培养2 d后，唾液干细胞球开始表达LN，培养第4 d后，随着唾
液干细胞球的增加，层连蛋白表达更明显。结论：应用该方法获得了唾液腺干细胞，具有成体干细胞的特征，具有自我更新和多向
分化的潜能。 |
| 英文摘要: |
| Objective:To develop an efficient method for isolation and culture of mouse salivary gland stem cells.Methods:Salivary gland stem cells were isolated and cultured from 7~8 week old C57BL/6J mice salivary gland. Inverted microscope was used to
observe the cells' morphology; Growth curve was determined by cell counting; the expression of laminin was analyzed by
Immunofluorescence.Results:After two days in culture, small aggregates of cells (salispheres) will be apparent in the cultures.
Salispheres will continue to grow in size over a period of four days in culture. The diameter is about 100 滋m and there are no obvious
spikes around the salispheres. After seven days in culture, obvious spikes occurred. The morphology of the salispheres after ten days was
different from those in four days; the salispheres have a trend to differentiation. Growth curve show the number of the salispheres was
increased in a time-dependent manner. At the same time, the diameter of the salispheres also increased significantly. These results
demonstrated that the salivary gland stem cells had high proliferation ability. Immunofluorescence results showed that salivary stem cells
began to express LN after 2 days in culture, and the level of LN was increased significantly after 4 days in culture.Conclusion:The
results of this study demonstrated that the mouse salivary gland stem cells from the mice salivary gland through this improved method
possessed the biological characteristics of adult stemcells. These cells had the ability to undergo both self - renewal and multi-directional
differentiation. |
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