Organotypic Cultures of Free-Floating Slices of Human Embryo Medulla Oblongata

Abstract

The aim of the present work was to produce organotypic cultures of human embryo brain tissues, as needed for investigation of the effects of serum factors on mental diseases and their pharmacological treatment. Cultures were made using slices of medulla oblongata from human embryos aged 9-10 weeks, obtained from medical abortions. Free-floating slices were cultured using a modified roller method for four weeks. Light and electron microscopy, along with immunocytochemistry, demonstrated that cultured slices showed differentiation and growth of neurons, astro- and microgliocytes, with formation of the synaptic contacts and glioneuronal interactions typical of organotypic cultures. However, along with differentiated neurons and glial cells, there was some persistence of undifferentiated cellular elements with signs of stem cells and neuron and gliocyte precursor cells. These experiments showed that organotypic roller cultivation of free-floating slices of human embryo brain tissue can be used to study the processes of neuron and gliocyte development, the mechanisms of neurotoxicity and neuroprotection using a variety of morphological (including stereological) and biochemical study methods.