Optimized isolation of renal plasma cells for flow cytometric analysis

Schaffer, Sandra; Maul-Pavicic, Andrea; Voll, Reinhard; Chevalier, Nina

doi:10.1016/j.jim.2019.06.019

Cited by 8 publications

(5 citation statements)

References 10 publications

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“…In addition, TACI harbors protease cleavage sites [ 1199 ] and can, therefore, be degraded or shedded when enzymes, e.g., collagenases, are used to dissociate tissues. For tissues such as the kidney, mild mechanical disruption in the absence of collagenase using the gentleMACS system (Miltenyi Biotech) is recommended [ 1200 ]. In tissues that strictly require enzymatic treatment (i.e., lamina propria of the gut), ENPP1 can be used in combination with CD138 to replace TACI.…”

Section: B Cell Phenotypesmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

et al. 2021

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The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.

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Section: B Cell Phenotypesmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

et al. 2021

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“…That is, the immune cells need to be released from the tissues by mechanical or enzymatic methods or combinations thereof. In particular, enzymatic digestion affects the detection of plasma cell markers, such as CD138 and CD267 (34). Therefore, we tested whether the newly identified ASC markers would resist to enzymatic digestion.…”

Section: Stable Expression Of Cd39 Cd81 Cd326 and Cd130 Allows Reliab...mentioning

confidence: 99%

CD39 and CD326 Are Bona Fide Markers of Murine and Human Plasma Cells and Identify a Bone Marrow Specific Plasma Cell Subpopulation in Lupus

et al. 2022

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Antibody-secreting cells (ASCs) contribute to immunity through production of antibodies and cytokines. Identification of specific markers of ASC would allow selective targeting of these cells in several disease contexts. Here, we performed an unbiased, large-scale protein screening, and identified twelve new molecules that are specifically expressed by murine ASCs. Expression of these markers, particularly CD39, CD81, CD130, and CD326, is stable and offers an improved resolution for ASC identification. We accessed their expression in germ-free conditions and in T cell deficient mice, showing that at least in part their expression is controlled by microbial- and T cell-derived signals. Further analysis of lupus mice revealed the presence of a subpopulation of LAG-3– plasma cells, co-expressing high amounts of CD39 and CD326 in the bone marrow. This population was IgM+ and correlated with IgM anti-dsDNA autoantibodies in sera. Importantly, we found that CD39, CD81, CD130, and CD326 are also expressed by human peripheral blood and bone marrow ASCs. Our data provide innovative insights into ASC biology and function in mice and human, and identify an intriguing BM specific CD39++CD326++ ASC subpopulation in autoimmunity.

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“…Therefore, a digestion protocol that optimizes enzyme concentration and time of digestion is needed to isolate cell types with minimal induction of apoptosis and aberrant gene expression. Recent publications have described digestion methods for specific tissues, including heart (6), brain (7), lung (8), kidney (9,10), and cultured cells (11,12), which have distinct extracellular matrix densities that require different enzymes and digestion times. Digestion cocktails can include enzymes such as trypsin, collagenase and elastase, or build on predesigned cocktails like Liberase (Sigma).…”

Section: Single Cell Isolationmentioning

confidence: 99%

Single Cell Analysis in Vascular Biology

Chavkin

Hirschi

2020

Front. Cardiovasc. Med.

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The ability to quantify DNA, RNA, and protein variations at the single cell level has revolutionized our understanding of cellular heterogeneity within tissues. Via such analyses, individual cells within populations previously thought to be homogeneous can now be delineated into specific subpopulations expressing unique sets of genes, enabling specialized functions. In vascular biology, studies using single cell RNA sequencing have revealed extensive heterogeneity among endothelial and mural cells even within the same vessel, key intermediate cell types that arise during blood and lymphatic vessel development, and cell-type specific responses to disease. Thus, emerging new single cell analysis techniques are enabling vascular biologists to elucidate mechanisms of vascular development, homeostasis, and disease that were previously not possible. In this review, we will provide an overview of single cell analysis methods and highlight recent advances in vascular biology made possible through single cell RNA sequencing.

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Optimized isolation of renal plasma cells for flow cytometric analysis

Cited by 8 publications

References 10 publications

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

CD39 and CD326 Are Bona Fide Markers of Murine and Human Plasma Cells and Identify a Bone Marrow Specific Plasma Cell Subpopulation in Lupus

Single Cell Analysis in Vascular Biology

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