CAJ | 학술논문

目的探讨两种神经球制片方法--贴片法和切片法的差异,并初探神经球内部的细胞结构.方法分离培养新生小鼠端脑神经干细胞,收集原代或传代培养7 d的神经球用于制片;贴片法是将神经球整体贴于载玻片上,切片法是将神经球用OCT包埋,冷冻切片机切片;通过比较两种制片方法的巢蛋白(nestin)免疫荧光染色的差异说明两种制片方法的差异;用HE染色方法将神经球切片染色,进一步观察其内部结构.结果成功分离培养的新生小鼠端脑神经干细胞在体外培养中形成神经球;贴片法制片神经球表面细胞染色良好,内部细胞不能着色,而且细胞形态显示不清晰,切片法制片神经球表面和内部细胞均能着色,细胞形态显示清晰:HE染色见神经球细胞之间借助突起相互连接,形成错综复杂的细胞网络.结论切片法制片较贴片法制片更有利于神经球的形态学研究;神经球是一个复杂的立体的细胞生长模式.
목적 탐토량충신경구제편방법--첩편법화절편법적차이,병초탐신경구내부적세포결구.방법 분리배양신생소서단뇌신경간세포,수집원대혹전대배양7 d적신경구용우제편;첩편법시장신경구정체첩우재파편상,절편법시장신경구용OCT포매,냉동절편궤절편;통과비교량충제편방법적소단백(nestin)면역형광염색적차이설명량충제편방법적차이;용HE염색방법장신경구절편염색,진일보관찰기내부결구.결과 성공분리배양적신생소서단뇌신경간세포재체외배양중형성신경구;첩편법제편신경구표면세포염색량호,내부세포불능착색,이차세포형태현시불청석,절편법제편신경구표면화내부세포균능착색,세포형태현시청석:HE염색견신경구세포지간차조돌기상호련접,형성착종복잡적세포망락.결론 절편법제편교첩편법제편경유리우신경구적형태학연구;신경구시일개복잡적입체적세포생장모식.
Objective To explore the differences of 2 slice techniques (paste-up method and section method) in the neurosphere slide preparation and the inside structure of the neurosphere. Methods Neural stem cells were isolated from the neonatal mouse telencephalon and the neurospheres on the 7~(th) d of primary culture or subculture were collected. Pasting the intact neurosphere to the slide was the paste-up method. Embedding the spheres with OCT followed by cutting into sections with a frozen section machine was the section method. Immunofluorescence was employed to observe the differences of nestin so as to illustrate the differences of the 2 neurosphere slide preparation methods. Inside structures of the neuroshperes were further studied by HE staining. Results Neural stem cells were successfully isolated and neurospheres formed during in vitro cultures. The superficial cells of the spheres were well stained, but the internal cells could not be stained when paste-up method was adopt; furthermore, the morphology of the cells could not be shown clearly. Both the superficial and internal cells could be well stained with clear morphology when the section method was chosed; as indicated by HE staining, the sphere cells connected to each other and formed complicated cell nets. Conclusion Section method is superior to paste-up method in morphology study of neurosphere and neurosphere enjoys a complicated three-dimension cell growth style.