FIGURE 8.

Post-thaw survival and function of cryopreserved D14 mSN under varying nucleation temperatures and cooling rates. Data are represented as mean ± 95% confidence interval. *p < 0.05; n.s.: p > 0.05. (A) Post-thaw recovery of D14 mSN showing no significant change with greater undercooling but trending significantly lower with faster cooling rate. n = 4. Red box: best-case parameters tested with the highest recovery. (B) Post-thaw reattachment of D14 mSN, normalized to reattachment of dissociated fresh cell control, showing no significant difference between any of the test conditions. n = 4. Crossed-out columns denoting insensitive post-thaw assay due to poor outcome of control cells. (C) Calcein AM-stained culture 24 and 48 h after replating dissociated (fresh) or cryopreserved (best-case) D14 mSN, showing a vast majority of live (calcein AM positive) cell population nonadherent or with rounded morphology in the first 24 h that was more sparsely distributed in the post-thaw culture, as well as subsequent nearly complete cell loss in both conditions at 48 h. Scale bar: 100 µm.