Comparison of two methods in neurosphere slide preparation
10.3760/cma.j.issn.1671-8925.2010.02.006
- VernacularTitle:两种神经球制片方法的比较研究
- Author:
Xin-Chun LIU
1
;
Yue ZHU
Author Information
1. 中国医科大学附属第一医院
- Keywords:
Neural stem cell;
Neurosphere;
Slide preparation;
Structure
- From:
Chinese Journal of Neuromedicine
2010;9(2):133-136
- CountryChina
- Language:Chinese
-
Abstract:
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.
