Experimental Studies On Culture Of Schwann Cells In Vitro

Peripheral nerve injury has many causes, such as trr :ma and tumor surgery. The regeneration and function recovery of peripheral nerve is one of the hottest issues in the field of neuroscience.The clinical treatment of a severed peripheral nerve involves the use of an autologous nerve graft to bridge the gap when it is difficult to suture it directly. But nerve autografting remains many problems,i.e.sacrificing one or more functional nerves and donor-site sequelaes include loss of sensation, scarring, etc. In addition, the limited availability of donor tissue represents a severe problem. In recent years, more attention has been focused on the development of tissure engineering nerv as a substitute for the nerve autograft to avoid these problems. The peripheral nerve tissue engineering technology brings new hope to resolve the difficulties in peripheral neuroscience.Objective: To find a new approach to obtain more population and more purification of SC from neonatal rat; Discuss the biological effects of basic fibroblast growth factor (b-FGF) to SC cultured in vitro.Method: Trypsin and high-density collagenase was used to separate SC from five to seven-day-old SD rat's sciatic nerves and brachial plexus. Cytosine arabinoside was used to eliminate fibroblast cells; then b-FGF was added to promote proliferation of SC. The growth and the biological characteristics of SC were observed by cell morphological survey, cell growth curve and S-100 stains.Result: SC separated from sciatic nerves and brachial plexus of SD rats can be cultured, purified and proliferated in vitro; b-FGF can promote the proliferation of SC; these SC can be identified with rabbit anti-S-100 protein antibody by SABC immunohistochemistry (IHC) methods. The percentage of SC purified was over 89%.Conclusion: 1. It is sure that Schwann Cells can be get from SD rat's sciaticnerves and brachial plexus.2. Ara-c can be used to purify Schwann cells cultured in vitro;b-FGF can promote the proliferation of SC. 3. Schwann Cells harvested from SD rats can be used as seeded cells in peripheral nerve tissue engineering.