Culture methods for selective growth of normal rat and human Schwann cells.

Beginning with an introduction of the Schwann cell itself, this chapter provides detailed methodology for growing pure Schwann cells of rat and human origin in serum-free medium without complicated cell purification schemes. Section II lists the essential preexperiment preparations such as materials, instruments, tissue culture medium, and solutions. Section III begins with the procedure for the dissection of embryonic dorsal root ganglia (DRG) from E14-E15 rat embryos, neonatal rats, and adult rats with diagrams; methods for enzymatic dissociation of the DRGs and the initiation of primary culture follow. A description of primary cultures with photographs is also provided for comparison. This section ends with a protocol for and results expected of serial passaging of rat Schwann cells. Section IV carefully describes the culture of Schwann cells from human adult nerve biopsy, a procedure that produces a thousandfold expansion of human Schwann cells within a month from initial plating. In the last section, basic immunocytochemistry as well as advanced in vitro remyelination techniques are provided for biochemical and functional characterization of Schwann cell cultures.