Cultures of cells from fetal rat brain: Methods to control composition, morphology, and biochemical activity

Fetal tissue transplantation is a promising new approach for the treatment of neurodegenerative diseases, but the optimal conditions for preparing cel ls for transplantation have not been defined. The growth of a population of septal brain cells, primarily containing cholinergic neurons and glia, was characterized after seeding at densities from 5 x 10(4) to 6 x 10(5) cells /cm(2), on polystyrene-, collagen-, laminin-, and fibronectin-coated surfac es, in the presence of serum and/or serum-free medium. Differentiated glial cells were selected by culture on fibronectin or laminin surfaces, in the presence of low amounts of serum (2.5% FBS) and G5, a soluble factor contai ning EGF and insulin. Differentiated neuronal cells were selected by cultur e on laminin, in the presence of low amounts of serum (2.5% FBS) and N2, a soluble factor containing supplemental hormones. In each case, a minimum se eding density of 1 x 10(5) cells/cm(2) was required. Neuronal growth could be maintained long term (21 days) with high levels of neuronal activity (Ch AT activity). (C) 1999 John Wiley & Sons, Inc.