Splenic B Cell Isolation
Splenic B Cell Isolation - Related Videos
17:07
Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS): Isolation of Splenic B Lymphocytes
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2023
Source: Perchet Thibaut1,2,3, Meunier Sylvain1,2,3, Sophie Novault4, Rachel Golub1,2,3 1 Unit for Lymphopoiesis, Department of Immunology, Pasteur Institute, Paris, France 2 INSERM U1223, Paris, France 3 Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France 4 Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, Center for Translational Science, Pasteur Institute, Paris, France The overall function of the immune system is to defend the body against infectious...
05:54
Induction of Graft Versus Host Disease in Mouse Models via Allogeneic Bone Marrow and Splenic Cell Transplantation
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2025
This video demonstrates the induction of intestinal graft-versus-host disease, GvHD, in a lethally-irradiated recipient mouse using allogeneic hematopoietic stem cells and GvHD-inducing splenic alloreactive T lymphocytes. This protocol helps establish a severe acute intestinal GvHD model to understand the mechanism of the disease and develop innovative therapeutic options.
09:09
An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets
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Cited by 8 •
2016
Distinct dendritic cell subsets exist as rare populations in lymphoid organs, and therefore are challenging to isolate in sufficient numbers and purity for immunological experiments. Here we describe a high efficiency, high yield method for isolation of all of the currently known major subsets of mouse splenic dendritic cells.
07:26
Isolation of Functional Cardiac Immune Cells
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Cited by 9 •
2011
This method for isolating functional immune cells from the heart provides an alternative to the conventional methods of collagenase digestion, which causes unwanted immune cell activation, resulting in a decreased responsiveness of these cells. Our method of isolation yields functional cardiac immune cells by avoiding problems associated with enzymatic digestion.
09:01
An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen
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Cited by 16 •
2015
Here we present an adapted protocol that can be used to generate a large number of murine invariant natural killer T cells from mouse spleen. The protocol outlines an approach by which splenic iNKT cells can be enriched for, isolated and expanded in vitro using a limited number of animals and reagents.
FAQs
Red blood cell lysis removes erythrocytes from the mechanically dissociated spleen preparation, producing a cleaner single-cell suspension for downstream enrichment and analysis. This step helps researchers examine B-cell populations without the spleen’s abundant red cell content obscuring measurements. The resulting preparation is better suited to flow cytometry, culture, molecular analysis, and comparisons between experimental groups.
Antibody-based magnetic separation uses antibodies to distinguish the desired B-cell population from other cells, allowing enrichment after tissue dissociation and red blood cell removal. Related sorting approaches also separate cells according to selected characteristics. The choice affects how specifically a defined B-cell population can be recovered for phenotype, function, activation, or molecular studies.
A defined population links measured responses more directly to B lymphocytes rather than to the mixed cells present in spleen tissue. Researchers can then assess phenotype and function, monitor activation, examine antigen presentation, and study antibody production under controlled conditions. This focused design also supports comparisons between healthy and disease models or between different immune challenges.
The workflow begins with mechanical dissociation of spleen tissue to create a single-cell suspension. Erythrocytes are then removed by red blood cell lysis, followed by enrichment through antibody-based magnetic separation or a related cell-sorting method. The recovered cells can proceed to flow cytometry, culture, molecular analysis, or functional experiments, depending on the research question.
Recovered B cells can be evaluated by flow cytometry to characterize phenotype, placed in culture to examine function and activation, or analyzed molecularly to investigate cellular responses. The isolated population also supports studies of antibody production and antigen presentation. Using several readouts allows researchers to connect B-cell identity with functional or immune-response outcomes.
Splenic B-cell preparations provide a defined population for comparing immune responses associated with infection or vaccination. Researchers can examine activation, antibody production, antigen presentation, and other functional changes in cells from relevant experimental groups. Comparisons with healthy or disease models help connect altered B-cell behavior to broader immunological responses.