Isolation, culture, and cryopreservation of adult rodent Schwann cells derived from immediately dissociated teased fibers

Natalia D. Andersen, Paula V Monje

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Adult Schwann cell (SC) cultures are usually derived from nerves subjected to a lengthy step of pre-degeneration to facilitate enzymatic digestion and recovery of viable cells. To overcome the need for pre-degeneration, we developed a method that allows the isolation of adult rat sciatic nerve SCs immediately after tissue harvesting. This method combines the advantages of implementing a rapid enzymatic dissociation of the nerve fibers and a straightforward separation of cells versus myelin that improves both cell yield and viability. Essentially, the method consists of (1) acute dissociation with collagenase and dispase immediately after removal of the epineurium layer and extensive teasing of the nerve fibers, (2) removal of myelin debris by selective attachment of the cells to a highly adhesive poly-l-lysine/laminin substrate, (3) expansion of the initial SC population in medium containing chemical mitogens, and (4) preparation of cryogenic stocks for transfer or delayed experimentation. This protocol allows for the procurement of homogeneous SC cultures deprived of myelin and fibroblast growth as soon as 3–4 days after nerve tissue dissection. SC cultures can be used as such for experimentation or subjected to consecutive rounds of expansion prior to use, purification, or cryopreservation.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages49-66
Number of pages18
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1739
ISSN (Print)1064-3745

Fingerprint

Schwann Cells
Cryopreservation
Rodentia
Myelin Sheath
Cell Culture Techniques
Nerve Fibers
Tissue and Organ Harvesting
Nerve Tissue
Cell Separation
Laminin
Collagenases
Sciatic Nerve
Peripheral Nerves
Mitogens
Adhesives
Lysine
Dissection
Digestion
Cell Survival
Fibroblasts

Keywords

  • Cryopreservation
  • Fibroblasts
  • Myelin
  • Peripheral nerve
  • Primary Schwann cell cultures
  • Teased fibers

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Andersen, N. D., & Monje, P. V. (2018). Isolation, culture, and cryopreservation of adult rodent Schwann cells derived from immediately dissociated teased fibers. In Methods in Molecular Biology (pp. 49-66). (Methods in Molecular Biology; Vol. 1739). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7649-2_4

Isolation, culture, and cryopreservation of adult rodent Schwann cells derived from immediately dissociated teased fibers. / Andersen, Natalia D.; Monje, Paula V.

Methods in Molecular Biology. Humana Press Inc., 2018. p. 49-66 (Methods in Molecular Biology; Vol. 1739).

Research output: Chapter in Book/Report/Conference proceedingChapter

Andersen, ND & Monje, PV 2018, Isolation, culture, and cryopreservation of adult rodent Schwann cells derived from immediately dissociated teased fibers. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1739, Humana Press Inc., pp. 49-66. https://doi.org/10.1007/978-1-4939-7649-2_4
Andersen, Natalia D. ; Monje, Paula V. / Isolation, culture, and cryopreservation of adult rodent Schwann cells derived from immediately dissociated teased fibers. Methods in Molecular Biology. Humana Press Inc., 2018. pp. 49-66 (Methods in Molecular Biology).
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