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. 2021 Dec 14;9:796960. doi: 10.3389/fcell.2021.796960

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Copyright © 2021 Li, Walsh, Truong, Petersen, Dutton and Hubel.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Post-thaw survival and function of cryopreserved D5 crest under varying nucleation temperatures and cooling rates. *p < 0.05; n.s.: p > 0.05. Red box: best-case parameters tested with the highest recovery and reattachment. (A) Post-thaw recovery of D5 crest showing no significant change with greater undercooling but trending significantly lower with faster cooling rate. n = 4. (B) Post-thaw reattachment of D5 crest showing similar trends as recovery but more distinct cell damage with cooling rates of −3°C/min and −5°C/min. n = 4. (C) Immunocytochemistry 48 h after re-culture of dissociated (fresh) or cryopreserved (best-case) D5 crest showing <1% TUJ1+, PRPN+ (white arrowheads) neuron differentiation, and a vast majority of resulted cell population with crest-like morphology and low-level, cytoplasmic TUJ1 and PRPN staining. Scale bar: 100 µm.