The methodological study of Schwann cell culture from sciatic nerve of adult SD rat

Objective Attempt to find out the method of isolated purified Schwann cells from sciatic nerve of adult SD rat Methods Three methods of culturing Schwann cells from sciatic nerve of adult rats were compared: (1) Conventional primary explant technique in tissue culture (Method A) (2) Conventional enzymatic disaggregating technique for cell culture (Method B) (3) A modified method of cell culture (Method C): Test group: we dissected very carefully and separated the sciatic nerve into individual fibers under surgical microscpe, individual fibers were cut into 0 5～1 mm pieces and treated with enzymatic disaggregating by twoenzymes (0 5% Trpsin and 0 06% Collagenase) in 37℃ for 80～90 min Control group: a application of conventional enzymatic disaggregating technique for culturing the nerve adventital tissues, which werethe remaining procedures of Test group We applied immunohistochemistry method with S100 antibody and Fibroblast antibody to identify Schwann cells Function of these Schwann cells proliferated were assessed by MTT assay and H 3 Thymidine incorporation assay. Conclusion In method C, a pure culture of Schwann cells was obtained, where the two techniques were used: careful separation of the sciatic nerve into individual fibers and use of higher concentrations of two enzymes for longer time to digest the nerve tissues These two technigues can cleanly remove and inhibit the fibroblasts, and enable healthy and normal proliferation of Schwann cells