CAJ | 학술논문

背景: 有关利用菲立磁体外细胞标记研究选择的动物主要是啮齿类动物,而研究灵长类动物食蟹猴的报道目前仍是空白.目的:观察利用菲立磁和转染试剂体外磁性标记食蟹猴骨髓基质细胞方法的可行性.方法:无菌条件下取食蟹猴骨髓,梯度密度离心法分离获取骨髓基质细胞,使用菲立磁-多聚赖氨酸复合物标记骨髓基质细胞,普鲁士蓝染色、电镜和锥虫蓝排除实验等方法鉴定菲立磁-多聚赖氨酸复合物标记食蟹猴骨髓基质细胞的效率和细胞的活力,倒置相差显微镜和免疫细胞化学方法检测骨髓基质细胞的增殖和分化能力.结果与结论:菲立磁可以高效率地标记骨髓基质细胞,标记效率在99%左右.光镜和电镜下骨髓基质细胞胞质内分别可见细小的蓝色铁颗粒和许多包裹铁颗粒的囊泡.菲立磁-多聚赖氨酸复合物标记对骨髓基质细胞的活力、增殖和分化等能力没有明显影响.提示菲立磁可以用来体外标记食蟹猴骨髓基质细胞.
배경: 유관이용비립자체외세포표기연구선택적동물주요시교치류동물,이연구령장류동물식해후적보도목전잉시공백.목적:관찰이용비립자화전염시제체외자성표기식해후골수기질세포방법적가행성.방법:무균조건하취식해후골수,제도밀도리심법분리획취골수기질세포,사용비립자-다취뢰안산복합물표기골수기질세포,보로사람염색、전경화추충람배제실험등방법감정비립자-다취뢰안산복합물표기식해후골수기질세포적효솔화세포적활력,도치상차현미경화면역세포화학방법검측골수기질세포적증식화분화능력.결과여결론:비립자가이고효솔지표기골수기질세포,표기효솔재99%좌우.광경화전경하골수기질세포포질내분별가견세소적람색철과립화허다포과철과립적낭포.비립자-다취뢰안산복합물표기대골수기질세포적활력、증식화분화등능력몰유명현영향.제시비립자가이용래체외표기식해후골수기질세포.
BACKGROUND: Studies regarding Feridex in vitro cell labeling are mainly in rodents, while little information is known on primate crab-eating macaque.OBJECTIVE: To explore the feasibility of protocols using Feridex and transfection agents for in vitro magnetic labeling of bone marrow stromal cells (BMSCs) in crab-eating macaque.METHODS: Under the sterile condition, the crab-eating macaque BMSCs were obtained by means of density gradient centrifugation following a bone puncture. Feridex-poly-l-lysine complexes were used to magnetically label BMSCs. The efficiency and cellular viability of Feridex-poly-l-lysine labeled BMSCs were evaluated by Prussian blue staining, electron microscopy, and trypan blue dye exclusion test. The proliferation and differentiation ability of Feridex-poly-l-lysine labeling BMSCs were also investigated by inverted phase contrast microscope and immunocytochemistry. RESULTS AND CONCLUSION: BMSCs could be effectively labeled by Feridex and labeling efficiency was around 99%. Tiny blue stained fine particles and numerous vesicles coated with the electron-dense magnetic iron particles could be found in the cytoplasm of Feridex-poly-l-lysine labeled BMSCs under optical microscopy and transmission electron microscopy respectively. Cell viability, proliferation and differentiation ability of labeled BMSCs were not affected by Feridex-poly-l-lysine labeling. Results indicated that Feridex might be used to label BMSCs of crab-eating macaque.