Rat Organotypic Cerebral Slice Culture And The Identification Of Pyramidal Cell

Objective: To establish a method of normal cerebral slice organotypic culture for rat, and identify the pyramidal cells in the organotypic cerebral slice. After organotypic brain slice culture is successful, we could establish various diseases of organotypic brain slice model on the basis of this.Method:Cerebral slice organotypic cultures were established successfully by using millicell-CM insert cultural technique and the culture solution composd of minimus culture media, blood serum of horse and Hank's liquor. Pyramidal cells survival in both organotypic cerebral slice and normal brain tissues in vivo were evaluated by culture morphology and nonphosphorylated neurofilament H (SMI-32) immunohistochemistry staining, according to the pyramidal cells in Cerebral cortex. To locate and compare in form the pyramidal cells both in Cerebral cortex and organotypic cerebral slice, To count and compare the numbers of the pyramidal cells in two weeks, three weeks and four weeks.Results:1 80% of the rat cerebral organotypic slice can survive in a month or more.2 In the cerebral cortex deep of cultivate brain slices, the cells of layers V are pyramidal and polygonal, diameter 25 to 50μm, with the uniform color of each cell bodies, shading deep in the cytoplasm. The dendritic of the cells are obvious, with a axon reach to cortex, which was very long. As cultivating the extension of time, the cells become more transverse protuberant growth.3 In the cerebral cortex in vivo the cells of layers V are pyramidal cells, of which the cell bodies present polygonal or elliptic, shading shallow of the cytoplasm, some of which show vacuolated. The dendritic are not obvious, but there is also a long axon reach to the cortex.4 The count number in two weeks, three weeks and four weeks were 28.50±4.34, 27.83±4.40, 27.67±3.55. Each datepoint of the cortex pyramidal cells count was no significant difference.Conclusion: Cerebral slice organotypic cultures were established successfully by using millicell-CM insert cultural technique and the culture solution composd of minimus culture media, blood serum of horse and Hanks'liquor, exploring and explicating the requirements of its growth process to the temperature, ph value, sugar content, nutrient-containing medium oxygen concentrations. Evaluating pyramidal cells of the organotypic cerebral slice by culture morphology and nonphosphorylated neurofilament H (SMI-32) immunohistochemistry staining can provide a solid scientific basis for oxygen-glucose deprivation organotypic cerebral slice, motor neuron disease, neurodegeneratie diseases and organotypic cerebral slice of traumatic brain diseases.