Using Flow Cytometry to Measure Phagocytic Uptake in Earthworms

Fuller-Espie, Sheryl L.

doi:10.1128/jmbe.v11i2.135

Cited by 8 publications

(3 citation statements)

References 8 publications

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“…) , rats , dogs , cats , cows , or cetaceans (Fig. ) , human dendritic cells , human peritoneal or monocyte‐derived macrophages , peritoneal‐ or bone‐marrow murine macrophages , and coelomocytes from earthworms . In addition, several established cell lines with phagocytic capacity can be used for experimental studies, typically the human monocytic cell lines U937 and THP‐1, or the murine macrophage cell lines J774A.1 and RAW 264.7 .…”

Section: Biological Applicationsmentioning

confidence: 99%

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

et al. 2019

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These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion.

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Section: Biological Applicationsmentioning

confidence: 99%

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

et al. 2019

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“…FCM assays of phagocytosis and other phagocytic‐related functions can be performed on a large variety of primary phagocytic cells, including but not restricted to peripheral blood monocytes and neutrophils from humans , rats , dogs , cats , cows or cetaceans , human dendritic cells , human peritoneal or monocyte‐derived macrophages , peritoneal‐ or bone‐marrow murine macrophages , and coelomocytes from earthworms . In addition, several established cell lines with phagocytic capacity can be used for experimental studies, typically the human monocytic cell lines U937 and THP‐1 or the murine macrophage cell lines J774A.1 and RAW 264.7 .…”

Section: Cytometric Parametersmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

et al. 2017

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International audienceThe classical model of hematopoiesis established in the mouse postulates that lymphoid cells originate from a founder population of common lymphoid progenitors. Here, using a modeling approach in humanized mice, we showed that human lymphoid development stemmed from distinct populations of CD127(-) and CD127(+) early lymphoid progenitors (ELPs). Combining molecular analyses with in vitro and in vivo functional assays, we demonstrated that CD127(-) and CD127(+) ELPs emerged independently from lympho-mono-dendritic progenitors, responded differently to Notch1 signals, underwent divergent modes of lineage restriction, and displayed both common and specific differentiation potentials. Whereas CD127(-) ELPs comprised precursors of T cells, marginal zone B cells, and natural killer (NK) and innate lymphoid cells (ILCs), CD127(+) ELPs supported production of all NK cell, ILC, and B cell populations but lacked T potential. On the basis of these results, we propose a "two-family" model of human lymphoid development that differs from the prevailing model of hematopoiesis

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“…15). Side scatter is a measure of cell granularity that is normally measured in particle uptake studies [101] and for our purposes, we looked at side scatter Since there are no reports in the literature on cell-particle assemblies involving irradiated cells, we had no indication as to whether to irradiate cells before or after hybrid assembly. We chose to irradiate cells subsequent to assembly since we wanted to avoid any possible change to cells induced by irradiation which may interfere with the ability to conjugate particles to the cell surface.…”

Section: Antibody-mediated Cell Surface Biotinylationmentioning

confidence: 99%

Assembly and characterization of a cell-particle hybrid system as a potential cancer vaccine

Ahmed¹

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Cancer vaccines represent a promising treatment modality for a worldwide health problem. Whether as an adjuvant or as a stand-alone therapy, cancer vaccines represent a tumor-specific and systemic treatment potentially capable of eliminating metastatic lesions without the severe side-effects often associated with chemotherapy. Specifically, whole cell tumor vaccines have shown promise in preclinical and clinical settings and the studies presented here represent the beginnings of an approach to improve the antitumor potency of these vaccines. This project demonstrates as "proof of concept" the feasibility of manufacturing tumor cell-particle hybrids. The coupled use of these two components, whole tumor cells and cargo-carrying biodegradable particles, as one entity in a cancer vaccine system is a new line of research. Stable cell-particle hybrids were assembled using avidin-biotin chemistry where cargo-carrying PLGA particles (500 nm diameter) were coated with streptavidin and allowed to bind to tumor cells that had been indirectly labeled with biotin (using an integrin-specific biotinylated antibody). That successful cell-particle hybrids were assembled was determined by multiple means, including flow cytometry, laser scanning confocal microscopy and scanning electron microscopy. Two murine tumor cell lines (representing melanoma and prostate cancer) were investigated in this study and successfully demonstrated the general applicability of the assembly method. Particles appeared to be localized on the cell surface (rather than endocytosed) as determined by microscopic imaging. The cell-particle hybrid was shown to be stable to irradiation, an important consideration since whole tumor cells need to be treated with ionizing radiation prior to being used as vaccines in order to render them nonproliferative and immunogenic. We also characterized loading and release profiles of CpG, a prospective vaccine adjuvant, into PLGA particles. v We conclude that we have developed a method for manufacturing cell-particle hybrids comprising PLGA nanoparticles and irradiated tumor cells. The next step would be to use CpG-loaded particles in the assembled hybrid and test the anti-tumor immune efficiency of this cancer vaccine formulation in either a melanoma or prostate cancer model. vi TABLE OF CONTENTS LIST OF TABLES …………………………………………………………….

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Using Flow Cytometry to Measure Phagocytic Uptake in Earthworms

Cited by 8 publications

References 8 publications

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

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

Guidelines for the use of flow cytometry and cell sorting in immunological studies^*

Assembly and characterization of a cell-particle hybrid system as a potential cancer vaccine

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Using Flow Cytometry to Measure Phagocytic Uptake in Earthworms

Cited by 8 publications

References 8 publications

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

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

Guidelines for the use of flow cytometry and cell sorting in immunological studies*

Assembly and characterization of a cell-particle hybrid system as a potential cancer vaccine

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Product

Resources

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Guidelines for the use of flow cytometry and cell sorting in immunological studies^*