dizocilpine-maleate and dibutyryl-cyclic-3--5--cytidine-monophosphate

We established selective primary cultures of neurons, astrocytes, and microglial cells from cryopreserved fetal cerebral cortex of cynomolgus monkeys (Macaca fascicularis). At 14 days in serum-containing medium, the cell cultures of the fetal cerebral cortex consisted primarily of neurons, astrocytes, and floating microglial cells. At 21 days, we observed a small number of myelin basic protein (MBP)-positive oligodendrocytes. The addition of cytosine arabinoside (a selective DNA synthesis inhibitor) at 2 days in culture eliminated proliferative glial cells, allowing adequate numbers of neurons to survive selectively. A chemically defined serum-free medium successfully supported neuronal survival at a level equivalent to that supported by the serum-containing medium. Brain-derived neurotrophic factor (BDNF) significantly affected the survival of primate neurons. Glutamate induced a significant degree of neuronal cell death against primate neurons and MK-801, a selective N-methyl-D-aspartate receptor (NMDAR) antagonist, blocked cell death, which suggests that primate cortical neurons have NMDAR and the glutamate-induced cell toxicity is mediated by NMDAR. In the serum-free medium, type-1 astrocytes responded to dibutyryl cyclic AMP and showed a process-bearing morphology. The growth of type-1 astrocytes in the serum-free medium was stimulated by epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and hydrocortisone, which are known growth factors in rat type-1 astrocytes. Cultured microglial cells expressed CD68, a monocyte marker. Macrophage-colony stimulating factor (M-CSF) stimulated microglial cell growth in the serum-free medium. These selective primary culture systems of primate cerebral cortical cells will be useful in issues involving species specificity in neuroscience.

Topics: Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Astrocytes; Cell Culture Techniques; Cell Death; Cell Division; Cells, Cultured; Cerebral Cortex; Culture Media, Serum-Free; Cyclic CMP; Cytarabine; Dizocilpine Maleate; Dose-Response Relationship, Drug; Epidermal Growth Factor; Excitatory Amino Acid Antagonists; Fetus; Fibroblast Growth Factors; Glial Fibrillary Acidic Protein; Glutamic Acid; Immunohistochemistry; Immunosuppressive Agents; Macaca fascicularis; Microglia; Microtubule-Associated Proteins; Myelin Basic Protein; Neurons