Culture of hypoglossal cells, dissociated from foetal and new-born rats.

Primary cultures of dissociated cells from brainstem cranial nuclei have not been described in the literature. The present paper shows that dissociated rat posterior brainstem cells, as well as cells from the hypoglossal nucleus, taken from both foetal and postnatal animals, can be maintained in long-term culture. This can be achieved by using a DMEM/F-12 medium with defined supplements, with or without foetal calf serum. Under such conditions, the growth of neuritic processes as well as the formation of neural networks can be observed. The different cell types present in the cultures can be identified by immunohistochemistry with antibodies raised against neuron specific enolase, glial fibrillary acidic protein, galactocerebroside and choline-acetyl transferase. As previously demonstrated for other brain-dissociated neurones maintained in cultures, the use of molecules, involved in cell adhesive mechanisms, can modify the morphological properties of the growing cells. This was particularly observed when poly-L-lysine and laminin were used as substrata.