Schwann cells are the primary inducers of regeneration of the peripheral nervous system. Schwann cells can be isolated from adult peripheral nerves, expanded in large numbers, and genetically transduced by viral vectors in vitro prior to their use in vivo. Here we describe how to use lentiviral vectors to transduce primary Schwann cells in vitro. We also describe how cultured Schwann cells can be used in conjunction with decellularized peripheral nerve sheaths prepared by multiple freeze thawing of peripheral nerve tissue. This process depletes all native cells from the nerve sheath but maintains basal lamina integrity and flexibility. A major advantage of using these decellularized nerve sheaths in repair strategies is that they can be obtained from cadaveric tissue and therefore do not require patient matching because the immune response is generated from the intrinsic cells and not the sheath itself. The patient's own cells can then be used to repopulate the decellularized peripheral nerve sheath. Our technique described in this chapter uses decellularized nerve sheaths which are repopulated with extrinsic Schwann cells previously grown in vitro. The Schwann cells can also be engineered in multiple ways, for example, to secrete bioactive proteins beneficial to axonal regeneration.
Keywords: Lentiviral vectors; Neurotrophins; Peripheral nerve; Regeneration; Schwann cells; Transduction.