Myelination competent conditionally immortalized mouse Schwann cells

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J Neurosci Methods. 2008 Jul 6.

Myelination competent conditionally immortalized mouse Schwann cells.

Saavedra JT, Wolterman RA, Baas F, Ten Asbroek AL.

Neurogenetics Laboratory, Academic Medical Center, P.O. Box 22700,

Amsterdam 1100 DE, The Netherlands.

Numerous mouse myelin mutants are available to analyze the biology of

the peripheral nervous system related to health and disease in vivo.

However, robust in vitro biochemical characterizations of players in

peripheral nerve processes are still not possible due to the limited

growth capacities of Schwann cells.

In order to generate cell lines from peripheral nerves that are

amenable to experimental manipulation, we have isolated Schwann cells

from transgenic mice (H-2Kb-tsA58) carrying the temperature sensitive

SV40 large T oncogene under the control of the interferon gamma

(IFNgamma) H-2Kb promoter.

These cells are immortalized at 33 degrees C when the SV40 large T

antigen has a stable conformation. At the non-permissive temperature

of 37 degrees C and in the absence of IFNgamma, the growth rate of

the cultures reduces and typical Schwann cell markers such as p75

(NGFR) become upregulated.

The conditionally immortalized Schwann cells allow genetic

manipulation as demonstrated here by the generation of a stable eGFP

expressing cell line. They regain their characteristic non-

immortalized properties at non-permissive temperature and

differentiate to myelin-forming cells when seeded on dorsal root

ganglia neurons.

The Schwann cell lines derived are valuable tools for in vitro

studies involving demyelinating diseases.