Cold storage of peripheral nerves: an in vitro assay of cell viability and function

The development of a nerve bank as a source of donor material to repair large defects in peripheral nerve injuries requires an understanding of the influence of cold storage on cell viability and function in these potential nerve grafts. Segments of peripheral nerves from both human and rat were stored in University of Wisconsin Cold Storage Solution (UW) at 4 degrees C for < 12 h, 3 days, and 1, 2, or 3 weeks. Cellular viability was initially assessed by the degree of cellular outgrowth from explants of the stored nerves placed in culture, and then further quantitated by dissociating the cultured nerve explants and calculating the type and number of cells per milligram of peripheral nerve. Rat Schwann cells (SCs) obtained from the stored (control and 1 and 2 weeks) nerves were tested for their functional ability to myelinate dorsal root ganglion (DRG) neurons in culture. Our findings indicate that human and rat peripheral nerves contain few viable SCs and fibroblasts after 3 weeks of cold storage with the quantity of viable cells within the human cold stored peripheral nerves decreasing significantly after 1 week of cold storage. Despite their reduced number, some SCs from rat nerves stored up to 2 weeks are capable of myelinating DRG axons in culture. These results suggest that short intervals (< 1 week) of cold storage will result in potential peripheral nerve grafts containing large populations of functional cells, while long-term (> or = 3 weeks) cold stored peripheral nerves will contain few viable cells.