A novel cryopreservation and biobanking strategy to study lymphoid tissue stromal cells in human disease
Joshua D Brandstadter1, Angelina De Martin2, Mechthild Lϋtge2
1Division of Hematology/Oncology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Insights
Cryopreservation enables banking of human lymphoid tissues for studying lymph node stromal cells (LNSCs) in disease. This method preserves cell viability and transcriptional profiles, facilitating LNSC research.
Area of Science:
- Immunology
- Cell Biology
- Histology
Background:
- Nonhematopoietic lymph node stromal cells (LNSCs) are crucial for immune responses and disease.
- Studying human LNSCs is challenging due to the need for fresh lymphoid tissue, often unavailable after diagnosis.
Purpose of the Study:
- To establish cryopreservation as a viable method for banking human lymphoid tissues for LNSC research.
- To assess the impact of cryopreservation on LNSC viability, cell type proportions, and transcriptional profiles.
Main Methods:
- Human tonsils and lymph nodes (LN) were cryopreserved.
- Enzymatic digestion was used to recover nonhematopoietic cells.
- Flow cytometry and single-cell transcriptomics were employed to analyze cell populations.
- In situ analyses confirmed cell distribution.
Main Results:
- Cryopreservation effectively preserved viable nonhematopoietic cells from lymphoid tissues.
- Comparable proportions of LNSC types were found in fresh and cryopreserved tissues.
- Transcriptional profiles remained largely unaffected by cryopreservation, showing overlap between tonsils and LN.
- Spatial distribution of cell types was maintained.
Conclusions:
- Cryopreservation is a suitable method for banking human lymphoid tissue for LNSC studies.
- This approach overcomes limitations of tissue availability, enabling broader research into LNSCs' roles in human diseases.
- The findings support the use of cryopreserved tissues for investigating LNSCs in immunity and pathology.
Abstract:
Nonhematopoietic lymph node stromal cells (LNSCs) regulate lymphocyte trafficking, survival, and function for key roles in host defense, autoimmunity, alloimmunity, and lymphoproliferative disorders. However, the 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 and lymph nodes (LN), lymphoid tissue fragments were cryopreserved for subsequent enzymatic digestion and recovery of viable nonhematopoietic cells. Flow cytometry and single-cell transcriptomics identified comparable proportions of LN stromal cell types in fresh and cryopreserved tissue. Moreover, cryopreservation had little effect on transcriptional profiles, which showed significant overlap between tonsils and LN. The presence and spatial distribution of transcriptionally defined cell types were confirmed by in situ analyses. Our broadly applicable approach promises to greatly enable research into the roles of LNSCs in human disease.
