Sciatic Nerve Dissection and Culturing

Adult male female Wistar rats were anaesthetised and sacrificed. The sciatic nerve was removed on both sides and transferred to a sterile work bench. The removal of epineurium and connective tissue was performed on ice. The nerves were kept in Hanks BSS (HBSS) + 1% (PAA Cölbe, Germany) Penicillin/Streptomycin (PAA, Cölbe, Germany) during preparation. Nerve fascicles were easily removed by pulling them out with sterile forceps. The fascicles were then cut into 3–4 mm pieces and transferred in 6-well dish (TPP, Trasadingen, Switzerland), followed by an incubation at 37 °C and 5% CO₂ with DMEM high glucose (Biochrome, Berlin, Germany), + 10% FCS (Biochrome, Berlin, Germany), + 1% P/S (PAA, Cölbe, Germany) and + 1% sodium pyruvate (Biochrome, Berlin, Germany) for 3 weeks. After 7 days, first migration of cells was observable [41 (link)].