A novel cryopreservation and biobanking strategy to study lymphoid tissue stromal cells in human disease
Insights
Cryopreservation enables banking of lymphoid tissue for studying non-hematopoietic lymph node stromal cells (LNSCs) in human diseases. This method preserves LNSC cell types and transcriptional profiles, facilitating crucial research.
Area of Science:
- Immunology
- Cell Biology
- Cryobiology
Background:
- Non-hematopoietic lymph node stromal cells (LNSCs) are crucial for immune responses but studying them in human diseases is challenging due to reliance on fresh tissue.
- Excised lymphoid tissues are often unavailable for LNSC research before diagnosis.
Approach:
- This study demonstrates cryopreservation of human tonsil tissue for LNSC isolation and analysis.
- Enzymatic digestion and recovery of viable non-hematopoietic cells were performed on cryopreserved samples.
- Flow cytometry and single-cell transcriptomics were used to analyze LNSC populations.
Key Points:
- Cryopreservation effectively preserves the proportions of LNSC cell types.
- Transcriptional profiles of LNSCs remain largely unchanged after cryopreservation.
- Cryopreserved tissue shows transcriptional overlap with lymph nodes, validating its utility.
- In situ analyses confirmed the presence and spatial distribution of identified cell types.
Conclusions:
- Cryopreservation is a viable method for banking lymphoid tissue for LNSC research.
- This approach overcomes limitations of studying LNSCs in human diseases.
- Enables broader research into LNSC roles in immunity, autoimmunity, and lymphoproliferative disorders.
Abstract:
Non-hematopoietic lymph node stromal cells (LNSCs) regulate lymphocyte trafficking, survival, and function for key roles in host defense, autoimmunity, alloimmunity, and lymphoproliferative disorders. However, study of LNSCs in human diseases is complicated by a dependence on viable lymphoid tissues, which are most often excised prior to establishment of a specific diagnosis. Here, we demonstrate that cryopreservation can be used to bank lymphoid tissue for the study of LNSCs in human disease. Using human tonsils, lymphoid tissue fragments were cryopreserved for subsequent enzymatic digestion and recovery of viable non-hematopoietic cells. Flow cytometry and single-cell transcriptomics identified comparable proportions of LNSC cell types in fresh and cryopreserved tissue. Moreover, cryopreservation had little effect on transcriptional profiles, which showed significant overlap between tonsils and lymph nodes. The presence and spatial distribution of transcriptionally defined cell types was confirmed by in situ analyses. Our broadly applicable approach promises to greatly enable research into the roles of LNSC in human disease.
