摘要
背景:目前尚缺乏高效、无创的方式将干细胞植入靶器官,探索引导干细胞到达靶器官或组织的途径以及提高干细胞归巢效率是现今干细胞研究的重点领域之一。目的:利用生物素-链霉亲和素反应体系建立一种简单可行的细胞表面化学修饰方法,并评价此方法进行骨髓间充质干细胞表面标记的效率及其对细胞生物学功能的影响。方法:全骨髓培养法得到第3代骨髓间充质干细胞,采用流式细胞仪鉴定;以磺化生物素-N-羟基琥珀酰亚胺、生物素、链霉亲和素将黏附分子配体唾液酸化的路易斯抗原装备到骨髓间充质干细胞表面;通过荧光显微镜评估骨髓间充质干细胞表面标记的效率,锥虫蓝染色法检测骨髓间充质干细胞的活性,CCK-8比色法检测骨髓间充质干细胞的增殖功能;成脂、成骨诱导检测骨髓间充质干细胞多分化功能。结果与结论:(1)全骨髓培养法培养2周,可得到第3代骨髓间充质干细胞,细胞表达CD90,CD29,不表达CD34和CD45。(2)以生物素及链霉亲和素成功将黏附分子配体唾液酸Lewis X(Sle X)装备到骨髓间充质干细胞表面,且对细胞活性、增殖、分化功能影响不大。(3)运用这种方法对细胞进行表型修饰,操作技术简单,修改效率可达88%,有望提高骨髓间充质干细胞的归巢率,未来会有广泛和重要的应用价值。
BACKGROUND： Currently, there is a lack of efficient, non-invasive way to transplant stem cells to the target organ or tissue. Exploring a way to guide targeting transplantation of stem cells and to improve the efficiency of stem cell homing is now one of focuses in the field of stem cells research. OBJECTIVE： To establish a simple and feasible method to chemically modify the cell surface using biotin-streptavidin reaction system, and to evaluate the efficiency of this method to label bone marrow mesenchymal stem cells（BMSCs） and its effects on cell biological functions. METHODS： Passage 3 BMSCs were obtained by whole bone marrow culture method and verified by flow cytometry. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were used to equip the adhesion molecule ligand, sialyated LewisX（SLeX）, to the BMSCs surface. The labeling rate of BMSCs was assessed using fluorescence microscope, the vitality of BMSCs was evaluated by trypan blue staining, and the proliferation of BMSCs was evaluated by cell counting kit-8 assay. Adipogenic and osteogenic inductions were used to evaluate the effect of the method on the multi-differentiation function of BMSCs. RESULTS AND CONCLUSION： After culture for 2 weeks, passage 3 BMSCs were obtained and confirmed by expressing CD90, CD29 and lack of CD34, CD45. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were successfully used to equip sialyated Lewis X（SLe X） to the BMSCs surface and had minor effects on the vitality, proliferation, and differentiation of BMSCs. This method was simple for surface modification and had a high modification rate of 88%. The homing of BMSCs modified by this method to the target organ or tissue could be greatly enhanced. Therefore, this method potentially could have extensive and important applications.
出处
《中国组织工程研究》
CAS
北大核心
2016年第10期1382-1388,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金资助项目(81260115)
江西省自然科学基金资助项目(20122BAB205005)~~
