[Organotypic cultures of free-floating sections of human embryonic medulla oblongata]

The aim of the present study was the development of organotypic culture of embryonic nerve tissue of human brain, which is necessary for the study of the effect of blood serum factors in mental diseases and their medicinal treatment. For the cultivation the sections of medulla oblongata were taken from human 9-10 week embryos obtained at legal medical abortion. Free-floating sections were cultured using a modified roller method for 4 weeks. By using the methods of light and electron microscopy, as well as of immunocytochemistry, it was demonstrated that within the cultured sections the differentiation and growth of neurons, astrocytes and microgliocytes took place accompanied by the formation of synaptic contacts and the establishment of glial-neuronal relationships, characteristic to the organotypical cultures. However, along with the differentiated neurons and glial cells, undifferentiated cellular elements were partially retained, that had the signs of neuronal and glial stem cells and progenitor cells. The studies performed have demonstrated that the method of organotypic roller culture of free-floating sections of human embryonic brain tissue could be applied for the study of the processes of neuronal and gliocyte development, mechanisms of neurotoxicity and neuroprotection using various morphological (including stereological) and biochemical research methods.