Characterization of human cultured astrocytes

Most studies on astrocytes are performed using cells from the central nervo us system of animals bred in laboratories. The recent knowledge in biology and molecular genetic shows that there are many metabolic and physiological differences between species and in the same species, differences exist bet ween strains. For this reason, it is preferable to work with human astrocyt es. The aim of the present work is to make available a simple method to eas ily get human astrocytes, even for studies necessitating a large number of cells. For this, cerebral cortices of human foetuses, more than 12 weeks of amenorrhoea- old, were got from the anatomo-pathology laboratory of a hosp ital. Explants were incubated in the Clang D medium and cells outgrew and a dhered to the flask. The explants were removed and the cell layers grew up to confluence. The cells were replated using DMEM medium up to fifteen pass ages. The shape of the cells was polygonal, thus looking like rodent astroc ytes. For the characterization purpose, the cells were immunostained and we re positive to anti-glial fibrillary acidic protein antibodies. A second ch aracterization was made by looking for glycogen particles in the cell cytop lasms by the means of electron microscopy. Beta and alpha particles were pr esent in the cells. Because glial fibrillary acidic protein is the specific marker of astrocytes and because the latter cells are those that accumulat e glycogen in the brain, it is concluded that almost all the cells are astr ocytes. From the preceding culture, many subcultures could be done. The sta ges of 16 and 22 weeks of amenorrhoea were the ages of the cells, which wer e successfully cultured, and no morphological difference was observed. The present research shows the possibility of getting a large number of human a strocytes for all type of study if the legislation of the country allows wo rking on human materials. So, this will minimalize the error due to an extr apolation from animal astrocytes to human astrocytes.