Age-related changes in attachment and proliferation of mouse Schwann cells in vitro

Schwann cells can be cultured readily from the peripheral nerves of the neonatal animal but not from the adult. To correlate the physiological properties of Schwann cells relevant to such a difference, we examined age-related changes in attachment and proliferation of mouse Schwann cells in vitro. The capacity of Schwann cells to attach to polylysine-coated coverslips at 1 day in vitro declined rapidly between 3 and 30 days of age, followed by a more gradual decrease with age. Attachment of Schwann cells from younger mice (but not older mice) was enhanced by precoating coverslips with laminin or to a lesser degree with fibronectin, suggesting an age-dependent decrease in receptors for these substrates. Indeed, the staining for fibronectin receptor could be demonstrated in vivo, and was more intense and diffuse in neonatal sciatic nerves. In vitro, although staining of Schwann cells and fibroblasts was clear, there was no age-related difference for the intensity or distribution of the staining. Proliferation, as assessed by thymidine incorporation at 1 day in vitro, was high when Schwann cells were isolated from younger mice but declined as a function of the age of mice from which cells were prepared. Removal of axonal and myelin debris from cultures 3 h after plating resulted in a reduction of thymidine uptake by Schwann cells from 30-day-old mice, but much less from 10-day-old mice. Schwann cell growth was faster in the cells from younger mice than older ones, thus leading to early confluency and cell-contact inhibition in the former. In addition, evidence is presented that in medium supplemented with fetal bovine serum, thymidine uptake by Schwann cells from mice at 3-30 days of age was three times higher than that by Schwann cells from age-matched rats. These results indicate that the methodology usually used for purification of rat Schwann cells involving antimitotics is not suitable for highly proliferating mouse Schwann cells.