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Science
Figure 1. Whole tissue cryopreservation preserves cell viability to allow biobanking of lymphoid tissue.
Figure 7. Tonsillar stromal cells have LN correlates and model LN tissue.
Figure 4. Whole tissue cryopreservation is feasible with different DMSO-containing reagents.
Figure 5. Cryopreservation of enzymatically digested cells impairs viability but preserves stromal cell subsets.
Figure 2. Single-cell RNA sequencing of CD45-EpCAM- cells identifies distinct stromal cell subsets in fresh lymphoid tissue.
Figure 8. Overlapping tonsillar and LN stromal cell subsets with more inflammatory immune-interacting CCL19+ TRC observed in LNs.
Figure 6. Histologic appearance of lymphoid stromal cell subsets identified by single-cell RNA sequencing.
Figure 3. Whole tissue cryopreservation preserves diverse stromal cell subsets detected by single-cell RNA sequencing.
Conclusions
We demonstrate whole-tissue cryopreservation can facilitate the study of LNSCs in human tissue.
• LNSC viability comparable to fresh tissue.
• LNSC subsets proportions comparable to fresh tissue.
• Comparable viability and LNSC subset proportions between commercial cryopreservative and 10% DMSO/90% fetal bovine serum.
• Improved viability compared with cryopreservation of single-cell suspension of enzymatically digested tissue
• FSC subsets can be identified
architecturally by fluorescent microscopy
• Tonsillar LNSCs have LN correlates