title: A new cell culture protocol for enrichment and genetic modification of adult canine Schwann cells suitable for peripheral nerve tissue engineering.
abstract: Easily applicable techniques are presented to obtain high numbers of enriched canine Schwann cells (cSC) in a short time-window. The potential of adult SC for tissue engineering of peripheral nerves and ex vivo gene therapy is obvious from physiological events taking place after peripheral nerve transection [Haastert, K., Grothe, C., 2007. Gene therapy in peripheral nerve reconstruction approaches. Curr. Gene Ther. 7, 221-228]. The presented techniques were modified from a protocol for cultivation and expansion of adult cSC by others [Pauls, J., Nolte, C., Forterre, F., Brunnberg, L., 2004. Cultivation and expansion of canine Schwann cells using reexplantation. Berl. Munch. Tierarztl. Wochenschr. 117, 341-352] and own experiences in rodent and human SC cultivation and transfection [Haastert, K., Mauritz, C., Chaturvedi, S., Grothe, C., 2007. Human and rat adult Schwann cell cultures: fast and efficient enrichment and highly effective non-viral transfection protocol. Nat. Protoc. 2, 99-104]. A purity of about 80% cSC achieved by immunopanning techniques and selective culture conditions is 2.5 fold higher as previously reported (Pauls et al., 2004). Additionally, highly enriched cSC populations are available in 3-4 weeks, only half the time period reported previously (Pauls et al., 2004). Furthermore, electroporation and genetic modification of cSC is reported for the first time.
authors.affiliation: Hannover Medical School, Institute of Neuroanatomy, OE 4140 Carl-Neuberg-Str. 1, D-30625 Hannover, Germany. Haastert.Kirsten@mh-hannover.de
authors.forename: K
authors.initials: K
authors.lastname: Haastert
authors.forename: P
authors.initials: P
authors.lastname: Seef
authors.forename: V M
authors.initials: VM
authors.lastname: Stein
authors.forename: A
authors.initials: A
authors.lastname: Tipold
authors.forename: C
authors.initials: C
authors.lastname: Grothe
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary*; D004285:Dogs; D008297:Male; D009417:Nerve Tissue / Q000166:cytology* / Q000502:physiology; D012583:Schwann Cells / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000662:veterinary*