Origin, maturation and recruitment of mast cell precursors (original) (raw)
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The in vitro differentiation of mast cells
The Journal of Cell Biology
When cells from lymph nodes or thoracic duct of mice hyperimmunized with protein antigens are cultivated on embryo monolayers in the presence of the antigen, numerous clones of mast cells appear. The histochemical and ultrastructural characteristics of the cells permit their identification as mast cells and distinguish them from the phagocytic histiocytes that usually arise in abundance in similar cultures from unimmunized mouse cells or from immunized mouse cells cultured in the absence of the antigen. Only a few colonies of mast cells appeared in the latter cultures. The basis for the induction of mast cell differentiation is not known.
Differentiation and proliferation of embryonic mast cells of the rat
The Journal of Cell Biology
Histochemical reactions and radioautography were used to investigate the sequence of mast cell development in rat embryos. Mast cells arise ubiquitously in and are confined to the loose connective tissue in the embryo. The alcian blue--safranin reaction distinguishes between weakly sulfated and strongly sulfated mucopolysaccharides by a shift from alcian blue to safranin staining. Based on this reaction and morphologic characteristics, four stages were identified. Stage I mast cells are lymphocyte-like cells with cytoplasmic granules which invariably stain blue with the alcian blue-safranin reaction. In Stage II cells the majority of granules are alcian blue-positive, but some safranin-positive granules have appeared. Stage III mast cells are distinguished by a majority of safranin-positive cytoplasmic granules; some alcian blue-posltive granules still remain. Stage IV cells contain only safranin-positive granules. Thymidine-H 3 uptake and identification of mitotic figures indicates that mast cells in Stages I and II comprise a mitotic pool while those in Stages III and IV are mitotically inactive. The pattern of $3~)4 incorporation and the sequence of appearance of histochemically identifiable mast cell constituents corroborates division of the proliferation and differentiation of embryonic mast cells into the four stages described above. The process of formation of mast cell granules is interpreted as reflecting the synthesis and accumulation of a heparin precursor in alcian blue positive granules followed by the synthesis and accumulation of highly N-sulfated heparin along with mast cell chymase and finally histamine in safranin-positive granules.
PloS one, 2016
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 ...
Mast cell clones: a model for the analysis of cellular maturation
1982
Cloned mouse mast cells resemble, by ultrastructure, immature mast cells observed in vivo. These mast cell clones can be grown in the absence of any other cells, facilitating direct investigations of their biochemistry and function. We find that cloned mast cells express plasma membrane receptors (Fc~R) that bind mouse IgE with an equilibrium constant (KA) similar to that of normal mouse peritoneal mast cells. In addition, cloned mast cells do not display detectable la antigens and cannot enhance Ig secretion when added to lymphocyte cultures or mediate natural killer lysis. In the presence of I mM sodium butyrate, cloned mast cells stop dividing and acquire abundant electron-dense cytoplasmic granules similar to those of mature mast cells. Their histamine content increases concomitant with cytoplasmic granule maturation and may exceed that of untreated mast cells by 50-fold. Unlike peritoneal mast cells, cloned mast cells incorporate 35S04 into chondroitin sulfates rather than heparin. 1-hese findings demonstrate that, unlike fully differentiated mouse peritoneal mast cells, cloned immature mouse mast cells contain no heparin and low levels of histamine. In addition, they establish that high-affinity Fc, R are expressed early in mast cell maturation, well before completion of cytoplasmic granule synthesis and mediator storage. We have previously reported methods to clone mast ceils with normal karyotypes from mouse hematopoietic tissue in vitro (30). Our cloned mast cells contain less histamine than normal mouse peritoneal mast cells and resemble immature mast cells by morphology. Others have described similar findings with uncloned cells (22, 32, 38, 42, 43, 51, 53). Although some mast cells synthesize heparin when fully differentiated (23, 54), certain mast cell tumors are devoid of heparin and incorporate 3BSO4 into chondroitin sulfate exclusively (18). Histochemical evidence suggests that normal immature connective tissue mast cells (6) and the mast cells in the intestinal lamina propria of rodents (25, 50) also may synthesize glycosaminoglycans other than heparin, but this notion has not been confirmed directly. In this report, we show that cloned mast cells closely resemble, by ultrastructure, immature mast cells found in vivo. Cloned mast cells incorporate 35804 preferentially into chondroitin sulfates, confirming histochemical evidence that immature mast cells contain little heparin. In addition, cloned mast cell proliferation, cytoplasmic granule synthesis, and mediator storage can be modulated in vitro, permitting direct analysis of mast cell maturation.
Ex Vivo and In Vitro Primary Mast Cells
Innate Immunity, 2008
Mast cells are cells of the innate immunity whose biological responses are markedly modulated by effector molecules of adaptive immunity i.e. antibodies. They thus contribute to antiinfectious defense, but also to antibody-dependent inflammatory responses. They are especially well known as inducers of the allergic reaction. They are widely distributed in most tissues, but in low numbers. They are not readily purified, and with a poor yield. For these reasons, means to generate large numbers of homogenous non-transformed mast cells have been developed. We describe here 1) fractionation methods suitable for purifying mouse or rat peritoneal mast cells and for purifying human mast cells of various origins, and 2) conditions for generating pure cultured mast cell populations from mouse, rat and human tissues.
The International Journal of Cell Cloning, 1987
Mast cells * Connective tissue-type mast cells Mucosal mast Interleukin 4 * Mutant mouse-Hematopoietic stem cell cells * Interleukin 3 Abstract. Information about the differentiation of mast cells has increased remarkably in the past ten years. This progress has resulted from the introduction of techniques which developed in other fields of experimental hematology. Once mast cells were recognized as a progeny of multipotential hematopoietic stem cells, their unique differentiation processes were clarified. Although most of the progeny of stem cells leave the hematopoietic tissue after maturation, undifferentiated precursors of mast cells leave the hematopoietic tissue. Morphologically, unidentifiable precursors migrate in the bloodstream, invade the connective tissues or the mucosa of the alimentary canal, proliferate, and differentiate into mast cells. Even after their morphological differentiation, some mast cells retain an extensive proliferative potential. There are at least two subpopulations of mast cells: a connective-tissue type and a mucosal type. Connective tissue-type and mucosal mast cells can be distinguished by histochemical, electron microscopical, biochemical and immunological criteria; however, these two types can interchange, and their phenotypes are determined by the anatomical microenvironment in which their final differentiation occurs. Although biochemical natures of the anatomical microenvironment are unknown, molecules that support proliferation and differentiation of mast cells in vitro have been characterized, i.e., interleukin 3 and interleukin 4. In the next ten years, increased information about the differentiation processes will probably induce further understanding of mast cell functions.
Murine mast cell lines as indicators of early events in mast cell and basophil development
European Journal of Immunology, 2000
To study early events in mast cell/basophil development, the phenotype of a panel of murine cell lines at various stages of differentiation was determined. Based on the expression on various mast cell-specific proteases and several additional hematopoietic differentiation markers, the cell lines CFTL-15 and MCP5/L were clearly identified as mast cells, although with a relatively immature phenotype. These two cell lines express the high-affinity IgE receptor § -chain, the mouse mast cell protease (MMCP)-5 and the carboxypeptidase A (CPA). Bone marrow-derived mast cells and the transplantable mast cell tumor MTC were shown to express the IgE receptor § -chain, MMCP-5 and CPA, as well as the mast cell tryptase MMCP-6 and the chymase MMCP-4, a protease expressed only during late stages of mast cell differentiation. These two cell types thus display a more mature mast cell phenotype. In contrast, the cell lines P815 and 32D cl3 did not express any mast cell differentiation markers. Interestingly, the IC-2 cell line was shown to express several markers for immature mast cells and in addition MMCP-8, a serine protease which may represent a marker for mouse basophils. By antibody staining, almost all IC-2 cells were shown to express MMCP-8. This indicates that individual cells may simultaneously express both mast cell and basophil markers. Moreover, these findings suggest that an early branch point in hematopoietic development where mast cells and basophils have a common precursor cell may exist.