Culturing Successfully Adult Schwann Cells from Peripheral Nerve of Rats

Shin, Mi Young; Roh, Young Nae; Woo, Ji Hyun; Kim, Young Hoon; Whang, Kyung Tai; Cho, Kyung Suck; Kim, Dal Soo

Abstract

PURPOSE: Schwann cells are difficult to isolate from adult mammalian peripheral nerves because of the abundant connective tissue and the highly differentiated state of the cells, particularly those involved in the formation of myelin. It has been shown that in vivo, both neurons and Schwann cells are conditioned by a nerve lesion, speeding up the Schwann cell proliferation and the neuronal regeneration. In this study with adult rat peripheral nerves, we report that a conditioning lesion increases Schwann cells and of cells which successfully attach to a tissue culture striatum.
METHODS
The study was done with Sprague-Dawley male rats(250-300 g). Their left sciatic nerves were exposed, severed at the sciatic notch and deflected. After 14 days, with 20 mm segments the nerves were excised from the distal stump of the conditioned sciatic nerves. Schwann cells were cultured in RPMI-1640 media. The identity of the cells was verified with antibody staining using S-100.
RESULTS
The lesion evoked a progressive and significant increase in the number of cells obtained as early as 48 hr of the plating, until day 12. Decreasing the duration of enzyme digestion to 3 and 4 hrs markely increased the number of attached Schwann cells. The peak numbers of attached Schwann cells was observed between day 12 and day 14 of the plating. Most attached Schwann cells had typical oval-shaped cell bodies, with prominent nuclei and bipolar cell extensions, resulting in overall spindle shapes.
CONCLUSION
Our findings indicate that a conditioning lesion enables us to isolate and culture adult Schwann cells successfully from the peripheral nerves of rats.