Isolation of skin-derived precursors (skps) and differentiation and enrichment of their schwann cell progeny
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ABSTRACT This protocol describes methods of isolating skin-derived precursors (SKPs) from rodent and human skin, and for generating and enriching Schwann cells from rodent SKPs. SKPs are
isolated as a population of non-adherent cells from the dermis that proliferate and self-renew as floating spheres in response to fibroblast growth factor 2 (FGF2) and epidermal growth
factor (EGF). Their differentiation into Schwann cells and subsequent enrichment of these differentiated progeny involves culturing SKPs as adherent cells in the absence of FGF2 and EGF, but
in the presence of neuregulins, and then mechanically isolating the Schwann cell colonies using cloning cylinders. Methods for expanding and characterizing these Schwann cells are provided.
Generation of primary SKPs takes approximately 2 weeks, while differentiation of Schwann cells requires an additional 4–6 weeks. Access through your institution Buy or subscribe This is a
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REFERENCES * Aguayo, A.J., Kasarjian, J., Skamene, E., Kongshavn, P. & Bray, G.M. Myelination of mouse axons by Schwann cells transplanted from normal and abnormal human nerves. _Nature_
268, 753–755 (1977). Article CAS Google Scholar * David, S. & Aguayo, A.J. Axonal regeneration after crush injury of rat central nervous system fibres innervating peripheral nerve
grafts. _J. Neurocytol._ 14, 1–12 (1985). Article CAS Google Scholar * Pearse, D.D. et al. Transplantation of Schwann cells and olfactory ensheathing glia after spinal cord injury: does
pretreatment with methylprednisolone and interleukin-10 enhance recovery? _J. Neurotrauma._ 21, 1223–1239 (2004). Article Google Scholar * Pearse, D.D. et al. cAMP and Schwann cells
promote axonal growth and functional recovery after spinal cord injury. _Nat. Med._ 10, 610–616 (2004). Article CAS Google Scholar * Xu, X.M., Chen, A., Guenard, V., Kleitman, N. &
Bunge, M.B. Bridging Schwann cell transplants promote axonal regeneration from both the rostral and caudal stumps of transected adult rat spinal cord. _J. Neurocytol._ 26, 1–16 (1997).
Article CAS Google Scholar * Xu, X.M., Zhang, S.X., Li, H., Aebischer, P. & Bunge, M.B. Regrowth of axons into the distal spinal cord through a Schwann-cell-seeded mini-channel
implanted into hemisected adult rat spinal cord. _Eur. J. Neurosci._ 11, 1723–1740 (1999). Article CAS Google Scholar * Pinzon, A., Calancie, B., Oudega, M. & Noga, B.R. Conduction of
impulses by axons regenerated in a Schwann cell graft in the transected adult rat thoracic spinal cord. _J. Neurosci. Res._ 64, 533–541 (2001). Article CAS Google Scholar * Paino, C.L.
& Bunge, M.B. Induction of axon growth into Schwann cell implants grafted into lesioned adult rat spinal cord. _Exp. Neurol._ 114, 254–257 (1991). Article CAS Google Scholar * Kocsis,
J.D., Akiyama, Y., Lankford, K.L. & Radtke, C. Cell transplantation of peripheral-myelin-forming cells to repair the injured spinal cord. _J. Rehabil. Res. Dev._ 39, 287–298 (2002).
PubMed Google Scholar * Keirstead, H.S., Morgan, S.V., Wilby, M.J. & Fawcett, J.W. Enhanced axonal regeneration following combined demyelination plus Schwann cell transplantation
therapy in the injured adult spinal cord. _Exp. Neurol._ 159, 225–236 (1999). Article CAS Google Scholar * Fouad, K. et al. Combining Schwann cell bridges and olfactory-ensheathing glia
grafts with chondroitinase promotes locomotor recovery after complete transection of the spinal cord. _J. Neurosci._ 25, 1169–1178 (2005). Article CAS Google Scholar * Barakat, D.J. et
al. Survival, integration, and axon growth support of glia transplanted into the chronically contused spinal cord. _Cell Transplant._ 14, 225–240 (2005). Article CAS Google Scholar * Li,
R. Culture methods for selective growth of normal rat and human Schwann cells. _Methods Cell Biol._ 57, 167–186 (1998). Article CAS Google Scholar * Mason, P.W., Attema, B.L. &
DeVries, G.H. Isolation and characterization of neonatal Schwann cells from cryopreserved rat sciatic nerves. _J. Neurosci. Res._ 31, 731–744 (1992). Article CAS Google Scholar * Oda, Y.,
Okada, Y., Katsuda, S., Ikeda, K. & Nakanishi, I. A simple method for the Schwann cell preparation from newborn rat sciatic nerves. _J. Neurosci. Methods_ 28, 163–169 (1989). Article
CAS Google Scholar * Peulve, P., Laquerriere, A., Paresy, M., Hemet, J. & Tadie, M. Establishment of adult rat Schwann cell cultures: effect of β-FGF, α-MSH, NGF, PDGF, and TGF-β on
cell cycle. _Exp. Cell Res._ 214, 543–550 (1994). Article CAS Google Scholar * Vroemen, M. & Weidner, N. Purification of Schwann cells by selection of p75 low affinity nerve growth
factor receptor expressing cells from adult peripheral nerve. _J. Neurosci. Methods_ 124, 135–143 (2003). Article CAS Google Scholar * Assouline, J.G., Bosch, E.P. & Lim, R.
Purification of rat Schwann cells from cultures of peripheral nerve: an immunoselective method using surfaces coated with anti-immunoglobulin antibodies. _Brain Res._ 277, 389–392 (1983).
Article CAS Google Scholar * Casella, G.T., Bunge, R.P. & Wood, P.M. Improved method for harvesting human Schwann cells from mature peripheral nerve and expansion _in vitro_. _Glia_
17, 327–338 (1996). Article CAS Google Scholar * Kreider, B.Q. et al. Enrichment of Schwann cell cultures from neonatal rat sciatic nerve by differential adhesion. _Brain Res._ 207,
433–444 (1981). Article CAS Google Scholar * Moretto, G., Kim, S.U., Shin, D.H., Pleasure, D.E. & Rizzuro, N. Long-term cultures of human adult Schwann cells isolated from autopsy
materials. _Acta Neuropathol. (Berl)_ 64, 15–21 (1984). Article CAS Google Scholar * Scarpini, E., Kreider, B.Q., Lisak, R.P. & Pleasure, D.E. Establishment of Schwann cell cultures
from adult rat peripheral nerves. _Exp. Neurol._ 102, 167–176 (1988). Article CAS Google Scholar * Morrissey, T.K., Kleitman, N. & Bunge, R.P. Isolation and functional
characterization of Schwann cells derived from adult peripheral nerve. _J. Neurosci._ 11, 2433–2442 (1991). Article CAS Google Scholar * Rutkowski, J.L., Tennekoon, G.I. &
McGillicuddy, J.E. Selective culture of mitotically active human Schwann cells from adult sural nerves. _Ann. Neurol._ 31, 580–586 (1992). Article CAS Google Scholar * Rutkowski, J.L.,
Kirk, C.J., Lerner, M.A. & Tennekoon, G.I. Purification and expansion of human Schwann cells _in vitro_. _Nat. Med._ 1, 80–83 (1995). Article CAS Google Scholar * Toma, J.G. et al.
Isolation of multipotent adult stem cells from the dermis of mammalian skin. _Nat. Cell Biol._ 3, 778–784 (2001). Article CAS Google Scholar * Fernandes, K.J.L. et al. A dermal niche for
multipotent adult skin-derived precursor cells. _Nat. Cell Biol._ 6, 1082–1093 (2004). Article CAS Google Scholar * Fernandes, K.J. et al. Analysis of the neurogenic potential of
multipotent skin-derived precursors. _Exp. Neurol._ 201, 32–48 (2006). Article Google Scholar * Toma, J.G., McKenzie, I.A., Bagli, D. & Miller, F.D. Isolation and characterization of
multipotent skin-derived precursors from human skin. _Stem Cells_ 23, 727–737 (2005). Article CAS Google Scholar * Joannides, A. et al. Efficient generation of neural precursors from
adult human skin: astrocytes promote neurogenesis from skin-derived stem cells. _Lancet_ 364, 172–178 (2004). Article CAS Google Scholar * McKenzie, I.A., Biernaskie, J., Toma, J.G.,
Midha, R. & Miller, F.D. Skin-derived precursors generate myelinating Schwann cells for the injured and dysmyelinated nervous system. _J. Neurosci._ 26, 6651–6660 (2006). Article CAS
Google Scholar Download references AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Developmental Biology Group, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada
Jeffrey A Biernaskie, Ian A McKenzie, Jean G Toma & Freda D Miller * Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada Freda D Miller *
Department of Physiology, University of Toronto, Toronto, Ontario, Canada Freda D Miller Authors * Jeffrey A Biernaskie View author publications You can also search for this author inPubMed
Google Scholar * Ian A McKenzie View author publications You can also search for this author inPubMed Google Scholar * Jean G Toma View author publications You can also search for this
author inPubMed Google Scholar * Freda D Miller View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Freda D Miller.
ETHICS DECLARATIONS COMPETING INTERESTS Dr. Freda Miller is a consultant for Aggregate Therapeutics, a biotechnology company who has licensed patents covering SKPs technology from McGill
University, who owns the technology. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Biernaskie, J., McKenzie, I., Toma, J. _et al._ Isolation of
skin-derived precursors (SKPs) and differentiation and enrichment of their Schwann cell progeny. _Nat Protoc_ 1, 2803–2812 (2006). https://doi.org/10.1038/nprot.2006.422 Download citation *
Published: 25 January 2007 * Issue Date: December 2006 * DOI: https://doi.org/10.1038/nprot.2006.422 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this
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