Growth of myenteric plexus explant cultures in a serum‐free, hormone‐supplemented culture medium

Abstract

Explant cultures of the myenteric plexus from beneath the taenia coli of newborn guinea‐pig caeca were grown on glass substrates in a serum‐free, hormone‐supplemented culture medium. The growth of enteric neurons, glial cells and fibroblasts and their interactions in this medium were studied over 3 weeks in vitro, and compared with those of cells cultured in the presence of foetal calf serum and in serum‐free medium in the absence of added hormones. Enteric neurons, glial cells and fibroblasts survived in the serum‐free, hormone‐supplemented medium, but this was dependent on the presence of the hormone supplements and also on a brief initial period of exposure to serum‐supplemented medium.The pattern of development of the cultures grown in serum‐free medium, however, differed markedly from that of cultures grown in serum‐supplemented medium: the numbers of enteric glial cells and fibroblasts were substantially reduced in the absence of serum; neurites grew extensively on the glass substrate rather than being restricted to a carpet of glial cells; and reaggregation of neurons and glial cells into compact groups resembling differentiated enteric ganglia did not occur.This ability to grow enteric neurons in a defined chemical environment, without the presence of unknown serum components, thus offers an opportunity for detailed study of the effects of growth and trophic factors on neuronal survival, neurite outgrowth and neurochemical differentiation in the entericnervous system, and also for study of the factors involved in enteric ganglion formation.