Culture Glial Cells Of Optic Never And Observe Fetures

Optic never is part of central never,it is composed of axon radiating from retinal ganglia cell and glial cells,it is easier to study because of its simple compenent.It is very fit model for people to study central never injury and transplantation of never.Glia cells of optic never include astrocytes and oligodendrocytes, unmature astrocytes obtaining from mice from embryo 8 days to postnal 4 days could improve never regeneration. However,optic never in optical canal suffer frequently from trauma,So studying the function of glial cell is one of the studying about optic never injury and never transplantation.Cultured glial cell is the basis of thoses studies.This study find approach to separation and purification astrocytes and oligodendrocytes which warrants a further investigation on optic never injury and transplantation. Objective: The optical nerve in optical canal is frequently used to study the optic nerve trauma,the function varies with the part of optic never.To get the optical nerve in optical canal of the mice in P2 period properly and primary culture the mixed glial cells and separate and purify the astrocytes and oligodendrocytes. To observe the features of glial cells and identify the Ill if cells. Methods:By microsurgery,To obtain the optic never in optical canal of mice in P2 period correctly via craiotomy or via the lateral wall of the orbit.Mixed glial cells were primary cultured with conditioned medium.TO Detachment selectively cells by shaking the cultures on an orbital shaker,microscopic examination was used to identify the cells in each preparation .Last,the astrocytes and oligodendrocytes were identified by immunohistochemisty.through ABC assay and Flow cytornetric,the results were analysesed. Result: The approach by the lateral wall of the orbit is complicated with the difficulty in identifying the anatomic landmark such as the apophysis of optical canal and the entrance plane of the orbit but cause little breeding.Another approach by craniotomy is ralatively less labourious and easy to find anatomic landmark such as optical chiasma,but cause more breeding.Astrocytes proliferate rapidly than oligodendrocytes, preparations appear >98%pure,The astrocytes were showed positive reaction to glial fibrilllary acidic protein (GFAP). Oligodendrocytesl is difficult to proliferate .The oligodendrocytes were showed postive to myelin basic protein ( MBP). Conclusion: The approach by craniotomy is an easy,quick and effective way to get the opitical nerve in optic canal of the mice in P2 period. Collectively,the results indicate that it is an easy and quick and reliable approach to separation and purification astroglial and oligodendroglial cell from primary mixed glial cells of optical never in P2 Iv periods by shaking. This method has no any apparent injury on the cell抯 morphology .This preparations should significantly aid in efforts to examine the biochemistry,physiology,and pharmacology of this class of optic never ,in particularly,aid in efforts to study the role of astrocytes after injury of optic never.