Schwann cells are adult cells that support the peripheral nervous system by myelinating axons. Nerves are harvested and the Schwann cells are isolated and propogated.
The nerves are harvested, rinsed 3 times in Liebovitz’s L15, stripped of epineurium, and dissected into individual fascicles. The fascicles are cut into 2-3-mm segments, placed in 100-mm dishes, kept in a humidified atmosphere with 5% CO2, and expanded. The medium (D10) is replaced 3 times per week with Dulbecco’s modified Eagle’s medium, fetal calf serum (10%), heregulin B1 (10 nM) and forskolin (1 µM). The fascicles are maintained submerged, but not attached, in a large volume of medium. Under these conditions, human schwann cells divide within the fascicles at a faster rate than fibroblasts.
After 2–4 weeks, the fascicles are enzymatically dissociated and the cells are washed twice in L15 and 10% FCS before plating at 100,000 cells/ml onto 100-mm culture dishes coated with ammoniated rat tail collagen in 10 ml expansion medium.
EMBRYONIC DEVELOPMENT & STEM CELL COMPENDIUM
