Glial development in primary cultures established from normal and X-irradiated neonatal spinal cord

The glial population of the lumbosacral spinal cord of the rat can be markedly depleted by exposure to ionizing radiation during the first postnatal week. Identification of specific cell populations which survive the exposure to radiation is difficult in situ; therefore, the present investigation used in vitro approaches to address issues related to specific phenotypes and maturational states of glia in cultures derived from non-irradiated (control) and irradiated (experimental) lumbosacral spinal cords of 3-day-old rats. Cultures were established from the spinal cords 2 to 4 hours following irradiation and were compared to cultures from non-irradiated, littermate controls. By 4 days in vitro (DIV) the numbers of cells in experimental cultures were profoundly reduced when compared to controls, and this reduction persisted through the termination of the study (8 DIV). In addition to reduction in numbers, astrocyte phenotypes were altered in experimental cultures, with greater proportions of the astrocyte population being constituted by the flat angular, large angular, and pancake types and a lesser proportion by stellate cells. The non-astrocytic cell types were dramatically reduced as evidenced by the paucity of oligodendrocytes immunoreactive for galactocerebroside and of small, non-process bearing cells binding the lectin, Griffonia (Bandeiraea) simplicifolia, a marker for microglia. Experimental cultures contained an increased incidence of binucleate astrocytes, an increase not restricted to a particular astrocyte phenotype. This study established the feasibility of utilizing this combined in vivo/in vitro approach in assessment of glial populations in immature spinal cords, and further investigations are in progress using this model.