Figure 3. Whole tissue cryopreservation preserves diverse stromal cell subsets detected by single-cell RNA sequencing.

Tonsil pieces were either kept fresh in FBS-containing RPMI at 4°C for two days vs. cryopreserved for two days or cryopreserved for two months. (A) Linear regression of gene expression between freshly processed and cryopreserved tonsil (with “cryo” defined as an average expression of two-day and two-month cryopreserved tissue). Pearson correlation with associated p-value listed in graph with genes showing increased transcript abundance in cryopreserved tissue highlighted in red and decreased abudandance highlighted in blue. (B) Colors in each bar define proportions of each subset within the entire sample with fibroblastic stromal cells (FSCs) highlighted in grey, blood endothelial cells (BECs) in yellow, lymphatic endothelial cells (LECs) in green, and otherwise unidentified cells (other) in red. (C) Colors in each bar define proportions of each Seurat-defined cluster within the entire sample. Cluster identities were determined by expression of known markers from analysis of freshly processed tonsil in Figure 2.