The Purified Cultivation And Identification And Oxygen Deprivated Cultivation Of Oligodendrocytes From Neonatal Rat Cerebrum

Objective To culture, purify and identify the astrocytes and oligodendrocytes from rat cerebral tissue. And to study the influence of oxygen and glucose deprivation on O2A progenitors.Methods Based on the different properties of cellular adhesions,developmental time-courses and growth pattern of astroglial cells and oligodendrocytes,astrocytes and oligodendrocytes were cultured and purified by twice orbital shaking , and identified by immmunofluorecence technique. O2A progenitors were cultured in oxygen deprivation combining with low glucose medium to mimic hypoxic injury. Electron microscope was used to study the ultrastructural changes of O2A progenitors.Results (1) Astrocytes and oligodendrocytes in mixed culture appeared better growth and differentiation than the respectively purified neuroglial, especially for the oligodendrocytes. O2A progenitors were A2B5-positive. The mature oligodendrocytes were Gal-positive. (2) Electron microscope showed a bundle of filament in the cytoplasma of O2A progenitors. The number of apoptotic O2A cells increased after oxygen deprivation combined with low glucose for 6 hours, and the apoptotic cells increased with time depending. Most cells undergo necrosis after culturing in oxygen deprivation combined with low glucose medium for 10 hours.Conclusions (1) High ratios of purified oligodendrocytes can be obtained by twice orbital shaking and proper culture medium. (2) Oxygen deprivation combined with low glucose on O2A cultivation can mimic the ischemic-hypoxic changes on O2A progenitor in vivo.