Schwann_Cells：schwann_cells.docVIP

Title: Isolation, purification and expansion of myelination-competent neonatal mouse Schwann Cells Running Title: Myelination-competent mouse Schwann Cells Authors: Henrika Honkanen1, Outi Lahti1, Marja Nissinen2, Riina M. Myllyl?2, Satu P?iv?l?inen1, Maria H. Alanne2, Sirkku Peltonen3, Juha Peltonen2 Anthony M. Heape1 Affiliations: 1 Myelin Group, Department of Anatomy Cell Biology, University of Oulu, Aapistie 7A, 90014 Oulu, Finland. 2 NF1 Research Team, Department of Anatomy Cell Biology, University of Oulu, Aapistie 7A, 90014 Oulu, Finland. 3 Cell-Cell Interaction Group, Department of Dermatology, University of Turku, Savitehtaankatu 1, 20521 Turku, Finland. Corresponding author: Doc. Anthony M. Heape, The Myelin Group, Department of Anatomy Cell Biology, P.O. Box 5000 (Aapistie 7A), 90014 University of Oulu, Finland. E-mail: Anthony.Heape@oulu.fi Phone: (+358)-8-5375197 Fax: (+358)-8-5375172 Funding This work was funded by the NeoBio Research Program of the Finnish National Technology Agency (Tekes), and a research grant from the Academy of Finland. AMH is a Research Fellow of the Academy of Finland. Abstract Most studies of PNS myelination using culture models are currently performed using dorsal root ganglion neurons (DRGN) and Schwann cells (SC) pre-purified from the rat. However, the potential of the model is severely compromised by the lack of rat myelin mutants, and the published protocols work very poorly with cells from mice, for which numerous myelin mutants are available. Here, we describe the isolation, purification and expansion of wild-type, myelination-competent SCs from the sciatic nerves of 4-day-old mouse pups. The protocol consistently provides ~95 %-pure mouse SC yields of 1.9 - 3.3 x 106 cells from the sciatic nerves of 12 to 15 four-day-old mouse pups, within 14 - 20 days of the isolation of the nerves. The proliferation rate of the cultured mouse SCs, expressed as “fold growth/week”, ranges from 2.7 to 4.30 under optimal conditions.