Fluorescent Reporter Mice for Nerve Guidance Conduit Assessment: A High‐Throughput in vivo Model

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Abstract
Objective: Use of cell culture and conventional in vivo mammalian models to assess nerve regeneration across guidance conduits is resource‐intensive. Herein we describe a high‐throughput platform utilizing transgenic mice for stain‐free axon visualization paired with rapid cryosection techniques for low‐cost screening of novel bioengineered nerve guidance conduit performance.Methods: Interposition repair of sciatic nerve transection in mice expressing yellow fluorescent protein in peripheral neurons (Thy1.2 YFP‐16) was performed with various bioengineered neural conduit compositions using a rapid sutureless entubulation technique under isoflurane anesthesia. Axonal ingrowth was assessed at 3 and 6 weeks using epifluorescent microscopy following cryosectioning.Results: Mean procedure time (incision‐to‐closure) was less than 2½ minutes. Direct operational costs of a 3‐week experiment was calculated at $21.47 per animal. Tissue processing steps were minimized to aldehyde fixation, cryoprotection and sectioning, and rapid fluorescent dye staining for conduit visualization. Fluorescent microscopy readily resolved robust axonal sprouting at 3 weeks, with clear elucidation of ingrowth‐permissive, semipermissive, or restrictive nerve guidance conduit environments.Conclusion: A rapid and cost‐efficient in vivo platform for screening of nerve guidance conduit performance has been described.Level of Evidence: NA. Laryngoscope, E392–E392, 2018
Funding Information
  • National Institutes of Health

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