Premium
Fluorescent Reporter Mice for Nerve Guidance Conduit Assessment: A High‐Throughput in vivo Model
Author(s) -
Mohan Suresh,
Hernández Iván Coto,
Wang Wenjin,
Yin Kaiyang,
Sundback Cathryn A.,
Wegst Ulrike G.K.,
Jowett Nate
Publication year - 2018
Publication title -
the laryngoscope
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.181
H-Index - 148
eISSN - 1531-4995
pISSN - 0023-852X
DOI - 10.1002/lary.27439
Subject(s) - in vivo , nerve guidance conduit , yellow fluorescent protein , axon , biomedical engineering , sciatic nerve , fluorescence microscope , fixation (population genetics) , green fluorescent protein , microbiology and biotechnology , anatomy , fluorescence , chemistry , biology , medicine , physics , quantum mechanics , biochemistry , gene
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