Primary dissociated cell culture of fetal rat central nervous tissue. II. Immunocytochemical and ultrastructural studies of myelinogenesis.

It is generally considered that the dissociated cell culture is a type of monolayer culture, and when this method is used with the central nervous system (CNS) cells, synapses may be formed but myelin is either not formed at all or formed in very small quantities. So we tried to establish a dissociated cell culture of the CNS which contains many myelinated axons. Cerebra from 18-day-old fetal rat were dissociated into single cells and plated onto poly-D-lysine-precoated coverslips at high cell density. With the progress of cell maturation, neuronal and non-neuronal cell processes intermingled and piled up on each other, forming 3-dimensional structures. We were able to observe not only many mature neurons and remarkable synapses but also many myelinated axons. Moreover direct connections of oligodendrocyte (ODC) somas to myelin sheaths were demonstrated either immunocytochemically or ultrastructurally. The formation of the 3-dimensional structure is considered to provide a favorable microenvironment for the intimate interaction of neurons and ODCs leading to the formation of many myelinated axons. So this culture system may provide a useful model for the investigation of the details in myelinogenesis under physiological conditions and demyelination or remyelination under pathological conditions such as multiple sclerosis and allied diseases.