Cryopreservation does not compromise all multi-lineage capability of DE aggregates. (a) qRT-PCR analysis of hepatic gene expression of thawed cells at day 14 of hepatic differentiation (n = 3). (b) qRT-PCR analysis of pancreatic gene expression of thawed cells at day 10 of pancreatic differentiation (n = 3–5). (c) qRT-PCR analysis of CDX2 expression of thawed cells at day 7 of intestinal differentiation (n = 3). (d) qRT-PCR analysis of NKX2.1 and p63 expression of thawed cells between days 10–13 of lung differentiation (n = 3–4). (e) Quantification of HNFα+ cells (n = 3), PDX1+ cells (n = 3–4), CDX2+ cells (n = 3), and NKX2.1+ cells (n = 3) from immunostaining, respectively. Each value of gene expression was first normalized to the reference gene(s) and then to day 0 undifferentiated cells. Values are represented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. All n values correspond to independent experiments.