STAR Protocols (Jun 2024)

Protocol to culture enteric glial cells from the submucosal and myenteric plexi of neonatal and juvenile pig colons

  • Madison L. Caldwell,
  • Caleb A. Cook,
  • Chloe L. Mariant,
  • Melissa Touvron,
  • Jack Odle,
  • Anthony T. Blikslager,
  • Amanda L. Ziegler,
  • Laurianne Van Landeghem

Journal volume & issue
Vol. 5, no. 2
p. 103057

Abstract

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Summary: Here, we present our protocol to culture enteric glial cells from the submucosal and myenteric plexus of neonatal and juvenile pig colons. We describe steps for colon isolation, microdissection, and enzymatic and mechanical dissociation. We include procedures for passaging and analyzing cell yield, freeze/thaw efficiency, and purity. This protocol allows for the generation of primary cultures of enteric glial cells from single-cell suspensions of microdissected layers of the colon wall and can be used to culture enteric glia from human colon specimens.For complete details on the use and execution of this protocol, please refer to Ziegler et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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