Human Schwann cells in vitro. I. Failure to differentiate and support neuronal health under co-culture conditions that promote full function of rodent cells.

Abstract

Schwann cells (SCs) play critical roles in regeneration after injury to the peripheral nervous system and can also induce axonal regeneration in the central nervous system. Transplantation of purified SCs into sites of neural injury in rodents has confirmed the remarkable ability of these cells to promote axonal regrowth, suggesting that human application of SC transplantation could be valuable. In this report, we have compared the functional capacities of SCs derived from adult human and rodent nerves by of SCs derived from adult human and rodent nerves by maintaining SCs from these two sources in culture with sensory neurons. We noted that techniques commonly in use for maintaining pure rat SC populations are not sufficient to sustain populations of human SCs free of fibroblasts. In these co-cultures, human SCs express a limited profile of characteristic behaviors and they proliferate more slowly than rat SCs in response to axonal contact. Slow SC proliferation, relative to that of contaminating fibroblasts, leads to a high proportion of fibroblasts in the cultures. After 3 to 4 weeks of co-culture with neurons, human SCs express extracellular matrix molecules, but only partially ensheathe axons, whereas rat SCs differentiate, form basal lamina, and ensheathe or myelinate axons. Co-culture of sensory neurons with human (but not rat) SC preparations (or conditioned medium therefrom) leads to a progressive neuronal atrophy characterized by shrinking neuronal cell bodies and a decrease in the density of the neurite network in the culture dish. As the divergent effects of human and rat SCs on neuronal health were also observed in co-cultures with human sensory neurons, these effects reflect differences between the rat and human-derived SC populations, rather than a species mismatch between SCs and neurons. The marked differences in behavior observed between rat and human SCs derived by the same methods requires further exploration if human-derived SCs are to be considered in the treatment of disease. In a companion article we report experiments that define culture conditions more effective in promoting human SC function in vitro.