Chapter 32 Isolation of Rat Peritoneal Mast Cells in High Yield and Purity

Cooper, P H; Stanworth, D.R.

doi:10.1016/s0091-679x(08)60496-3

Cited by 14 publications

(8 citation statements)

References 12 publications

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“…These cells can be obtained by culturing bone marrow progenitors in the presence of interleukin (IL)-3 and SCF (Razin et al, 1981; Tsai et al, 1991). In rat, mast cells are often isolated in mature stage from peritoneal cavity (Cooper and Stanworth, 1976; Poole and Zetter, 1983; Gruber et al, 1994; Brzezińska-Błaszczyk et al, 2007). Human mast cells are usually obtained by culturing mast cell progenitors from various tissues in the presence of SCF, IL-6, and IL-10.…”

Section: Methods Used For Studies Of Mast Cell Chemotaxismentioning

confidence: 99%

Mast Cell Chemotaxis – Chemoattractants and Signaling Pathways

Hálová¹,

Dráberová²,

Dráber³

2012

Front. Immun.

154

150

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Migration of mast cells is essential for their recruitment within target tissues where they play an important role in innate and adaptive immune responses. These processes rely on the ability of mast cells to recognize appropriate chemotactic stimuli and react to them by a chemotactic response. Another level of intercellular communication is attained by production of chemoattractants by activated mast cells, which results in accumulation of mast cells and other hematopoietic cells at the sites of inflammation. Mast cells express numerous surface receptors for various ligands with properties of potent chemoattractants. They include the stem cell factor (SCF) recognized by c-Kit, antigen, which binds to immunoglobulin E (IgE) anchored to the high affinity IgE receptor (FcεRI), highly cytokinergic (HC) IgE recognized by FcεRI, lipid mediator sphingosine-1-phosphate (S1P), which binds to G protein-coupled receptors (GPCRs). Other large groups of chemoattractants are eicosanoids [prostaglandin E2 and D2, leukotriene (LT) B4, LTD4, and LTC4, and others] and chemokines (CC, CXC, C, and CX3C), which also bind to various GPCRs. Further noteworthy chemoattractants are isoforms of transforming growth factor (TGF) β1–3, which are sensitively recognized by TGF-β serine/threonine type I and II β receptors, adenosine, C1q, C3a, and C5a components of the complement, 5-hydroxytryptamine, neuroendocrine peptide catestatin, tumor necrosis factor-α, and others. Here we discuss the major types of chemoattractants recognized by mast cells, their target receptors, as well as signaling pathways they utilize. We also briefly deal with methods used for studies of mast cell chemotaxis and with ways of how these studies profited from the results obtained in other cellular systems.

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Section: Methods Used For Studies Of Mast Cell Chemotaxismentioning

confidence: 99%

Mast Cell Chemotaxis – Chemoattractants and Signaling Pathways

Hálová¹,

Dráberová²,

Dráber³

2012

Front. Immun.

154

150

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“…After 15-30 days, a cell suspension containing 2-5 % mast cells was obtained by peritoneal lavage with iso-osmotic saline (0.9% NaCI) containing heparin (25i.u./ml). The mast cells were purified to about 95% homogeneity by using the method of Cooper & Stanworth (1976) with minor modifications. We obtained about 106 cells/animal; four to six animals were used for each experiment.…”

Section: Methodsmentioning

confidence: 99%

Evidence for a role of phosphatidylinositol turnover in stimulus–secretion coupling. Studies with rat peritoneal mast cells

Cockcroft

Gomperts

1979

Biochemical Journal

104

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Histamine secretion and phosphatidylinositol turnover were compared in antigen-sensitized rat peritoneal mast cells stimulated with a number of different ligands. A small and variable increase in the incorporation of [(32)P]P(i) and of [(3)H]inositol into phosphatidylinositol was observed when the cells were treated with immunoglobulin E-directed ligands (antigens and concanavalin A), and this was accompanied by a low amount of secretion (<10% of total cell histamine). In the presence of added phosphatidylserine, the addition of immunoglobulin E-directed ligands invariably led to an enhanced rate (approx. 4-fold) of labelling of phosphatidylinositol and, in the presence of Ca(2+), this was accompanied by the secretion of histamine. The labelling of phosphatidylinositol and histamine secretion were also stimulated by chymotrypsin and compound 48/80. Whereas the phosphatidylinositol response did not require the presence of extracellular Ca(2+), the secretion of histamine was either enhanced or dependent on extracellular Ca(2+) (depending on the ligand used). The dependence on ligand concentration for the phosphatidylinositol response and histamine secretion were similar. The increased isotopic incorporation into phosphatidylinositol continued for about 1h although histamine secretion (elicited with concanavalin A) stopped within 2min. These results support the proposition that metabolic events involving phosphatidylinositol play a necessary intermediate role in the regulation of Ca(2+) channels by ligand-activated receptors.

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“…Also the isolation of MCs from tissue by enzymatic digestion protocols using collagenase and elastase resulted in cells with variable viability and purity [ 16 ]. Other protocols allowed the isolation of rat peritoneal MCs (PMCs) from a single peritoneal lavage resulting in high yield of MCs [ 17 ]. In addition, time consuming multistep separation techniques that combined enzymatic treatment, elutriation, magnetic CD14 + cell depletion (monocytes, macrophages) and cell sorting techniques with monoclonal antibodies directed against CD117/KIT were suitable to isolate and enrich MCs from human lung, skin, and uterus [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

Isolation of Mature (Peritoneum-Derived) Mast Cells and Immature (Bone Marrow-Derived) Mast Cell Precursors from Mice

et al. 2016

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Mast cells (MCs) are a versatile cell type playing key roles in tissue morphogenesis and host defence against bacteria and parasites. Furthermore, they can enhance immunological danger signals and are implicated in inflammatory disorders like fibrosis. This granulated cell type originates from the myeloid lineage and has similarities to basophilic granulocytes, both containing large quantities of histamine and heparin. Immature murine mast cells mature in their destination tissue and adopt either the connective tissue (CTMC) or mucosal (MMC) type. Some effector functions are executed by activation/degranulation of MCs which lead to secretion of a typical set of MC proteases (MCPT) and of the preformed or newly synthesized mediators from its granules into the local microenvironment. Due to the potential accumulation of mutations in key signalling pathway components of corresponding MC cell-lines, primary cultured MCs are an attractive mean to study general features of MC biology and aspects of MC functions relevant to human disease. Here, we describe a simple protocol for the simultaneous isolation of mature CTMC-like murine MCs from the peritoneum (PMCs) and immature MC precursors from the bone marrow (BM). The latter are differentiated in vitro to yield BM-derived MCs (BMMC). These cells display the typical morphological and phenotypic features of MCs, express the typical MC surface markers, and can be propagated and kept in culture for several weeks. The provided protocol allows simple amplification of large quantities of homogenous, non-transformed MCs from the peritoneum and bone marrow-derived mast cells for cell- and tissue-based biomedical research.

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Chapter 32 Isolation of Rat Peritoneal Mast Cells in High Yield and Purity

Cited by 14 publications

References 12 publications

Mast Cell Chemotaxis – Chemoattractants and Signaling Pathways

Mast Cell Chemotaxis – Chemoattractants and Signaling Pathways

Evidence for a role of phosphatidylinositol turnover in stimulus–secretion coupling. Studies with rat peritoneal mast cells

Isolation of Mature (Peritoneum-Derived) Mast Cells and Immature (Bone Marrow-Derived) Mast Cell Precursors from Mice

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