Description and Characterization of Three-Dimensional Human Mast Cell Progenitor Spheroids In Vitro

Human mast cells (MC) are an essential component of the immune system as they uniquely store and release a wide range of soluble mediators through IgE and non-IgE mechanisms. Several tissue sources can be used to differentiate functional MC for in vitro and in vivo studies. Here we describe an improved method for obtaining large numbers of human MC from adipose tissue with advantages over current methods. We analyzed donor parameters (e.g. age, race) on MC-isolation following adipose and skin tissue digestion from healthy donors. Adipose and skin-derived MC were morphologically and immunophenotypically similar in all donors regardless of age. However, donor-dependent variations in MC numbers were observed following tissue digestion. In addition, we identified and characterized three-dimensional structures from which mature MC emerged in vitro using peripheral blood and human tissue sources. MC progenitor spheroids (MCPS) appeared approximately one week following progenitor isolation and were consistently observed to have mature MC attached, emerging, or nearby when cultured in a stem cell factor-containing medium. The overall characteristics of the MCPS were similar from each tissue source. We propose that these MCPS serve as the common source of human MC in vitro.

[1]  M. Moonim,et al.  Update on diagnostic approaches and therapeutic strategies in systemic mastocytosis. , 2022, Best practice & research. Clinical haematology.

[2]  Sung Hyun Lee,et al.  Human Tumor Targeted Cytotoxic Mast Cells for Cancer Immunotherapy , 2022, Frontiers in Oncology.

[3]  C. Kepley,et al.  Harnessing the Anti-Tumor Mediators in Mast Cells as a New Strategy for Adoptive Cell Transfer for Cancer , 2022, Frontiers in Oncology.

[4]  C. Thiele,et al.  Generation of immune cell containing adipose organoids for in vitro analysis of immune metabolism , 2020, Scientific Reports.

[5]  L. Hellman,et al.  How Relevant Are Bone Marrow-Derived Mast Cells (BMMCs) as Models for Tissue Mast Cells? A Comparative Transcriptome Analysis of BMMCs and Peritoneal Mast Cells , 2020, Cells.

[6]  Sung Hyun Lee,et al.  A New Cellular Cancer Immunotherapy Platform Technology Using Autologous Human Adipose-Derived Mast Cells , 2020, Journal of Immunology.

[7]  G. Bandara,et al.  Description and Characterization of a Novel Human Mast Cell Line for Scientific Study , 2019, International journal of molecular sciences.

[8]  M. Penichet,et al.  Human Mast Cells From Adipose Tissue Target and Induce Apoptosis of Breast Cancer Cells , 2019, Front. Immunol..

[9]  B. Cousin,et al.  Rapid and Efficient Production of Human Functional Mast Cells through a Three-Dimensional Culture of Adipose Tissue–Derived Stromal Vascular Cells , 2018, The Journal of Immunology.

[10]  Joakim S. Dahlin,et al.  An Optimized Protocol for the Isolation and Functional Analysis of Human Lung Mast Cells , 2018, Front. Immunol..

[11]  Joakim S. Dahlin,et al.  Deciphering the differentiation trajectory from hematopoietic stem cells to mast cells. , 2018, Blood advances.

[12]  M. Tsai,et al.  Mast cells as sources of cytokines, chemokines, and growth factors , 2018, Immunological reviews.

[13]  D. Metcalfe,et al.  An optimized protocol for the generation and functional analysis of human mast cells from CD34+ enriched cell populations. , 2017, JIM - Journal of Immunological Methods.

[14]  D. Ribatti.  The development of human mast cells. An historical reappraisal. , 2016, Experimental cell research.

[15]  J. Wood,et al.  Mast Cell: A Multi-Functional Master Cell , 2016, Front. Immunol..

[16]  M. Moonim.  The bone marrow in systemic mastocytosis – an update , 2015 .

[17]  D. Ribatti,et al.  Tryptase, a novel angiogenic factor stored in mast cell granules. , 2015, Experimental cell research.

[18]  C. Oliver,et al.  Mast Cell Function , 2014, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[19]  T. Rülicke,et al.  A new human mast cell line expressing a functional IgE receptor converts to tumorigenic growth by KIT D816V transfection. , 2014, Blood.

[20]  E. Crivellato,et al.  Mast cell ontogeny: an historical overview. , 2014, Immunology letters.

[21]  P. Kovanen,et al.  Human mast cells arise from a common circulating progenitor. , 2013, The Journal of allergy and clinical immunology.

[22]  P. Bruhns.  Properties of mouse and human IgG receptors and their contribution to disease models. , 2012, Blood.

[23]  K. Austen,et al.  Protease phenotype of constitutive connective tissue and of induced mucosal mast cells in mice is regulated by the tissue , 2011, Proceedings of the National Academy of Sciences.

[24]  R. Korthuis,et al.  Mast Cell Proteases and Inflammation. , 2011, Drug discovery today. Disease models.

[25]  J. Steinke,et al.  Characterization of a novel human mast cell line that responds to stem cell factor and expresses functional FcεRI. , 2011, The Journal of allergy and clinical immunology.

[26]  B. Cousin,et al.  Adipose Tissue as a Dedicated Reservoir of Functional Mast Cell Progenitors , 2010, Stem cells.

[27]  R. Lenk,et al.  A New Class of Human Mast Cell and Peripheral Blood Basophil Stabilizers that Differentially Control Allergic Mediator Release , 2010, Clinical and translational science.

[28]  T. Moon,et al.  Advances in mast cell biology: new understanding of heterogeneity and function , 2010, Mucosal Immunology.

[29]  H. Horny.  Mastocytosis: an unusual clonal disorder of bone marrow-derived hematopoietic progenitor cells. , 2009, American journal of clinical pathology.

[30]  S. Bischoff.  Role of mast cells in allergic and non-allergic immune responses: comparison of human and murine data , 2007, Nature Reviews Immunology.

[31]  Shoko Iwaki,et al.  Generation, Isolation, and Maintenance of Rodent Mast Cells and Mast Cell Lines , 2006, Current protocols in immunology.

[32]  Wei Zhao,et al.  FcγRIIa, Not FcγRIIb, Is Constitutively and Functionally Expressed on Skin-Derived Human Mast Cells1 , 2006, The Journal of Immunology.

[33]  M. Gershwin,et al.  Mastocytosis , 2006, Clinical reviews in allergy & immunology.

[34]  T. Suga,et al.  Detection of tryptase−, chymase+ cells in human CD34+ bone marrow progenitors , 2004, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[35]  C. Hughes,et al.  Of Mice and Not Men: Differences between Mouse and Human Immunology , 2004, The Journal of Immunology.

[36]  D. Metcalfe,et al.  Characterization of novel stem cell factor responsive human mast cell lines LAD 1 and 2 established from a patient with mast cell sarcoma/leukemia; activation following aggregation of FcepsilonRI or FcgammaRI. , 2003, Leukemia research.

[37]  C. Kepley,et al.  Evidence for human mast cell nonreleaser phenotype. , 2003, Journal of Allergy and Clinical Immunology.

[38]  Y. Shimizu,et al.  Somatic mutations in the p53 gene account for the extension of replicative life span of macaque cells. , 2002, Biochemical and biophysical research communications.

[39]  J. Kochan,et al.  Human skin-derived mast cells can proliferate while retaining their characteristic functional and protease phenotypes. , 2001, Blood.

[40]  H. W. Chang,et al.  An improved procedure for the development of human mast cells from dispersed fetal liver cells in serum-free culture medium. , 2000, Journal of immunological methods.

[41]  B. Foster,et al.  Demonstration that human mast cells arise from a progenitor cell population that is CD34(+), c-kit(+), and expresses aminopeptidase N (CD13). , 1999, Blood.

[42]  M. Triggiani,et al.  Molecular and cellular biology of mast cells and basophils. , 1997, International archives of allergy and immunology.

[43]  C. Kepley,et al.  Quantitation of tryptase, chymase, Fc epsilon RI alpha, and Fc epsilon RI gamma mRNAs in human mast cells and basophils by competitive reverse transcription-polymerase chain reaction. , 1995, Journal of immunology.

[44]  G. Marone,et al.  Human heart mast cells: a definitive case of mast cell heterogeneity. , 1995, International archives of allergy and immunology.

[45]  E. Arbustini,et al.  Human heart mast cells. Isolation, purification, ultrastructure, and immunologic characterization. , 1995, Journal of immunology.

[46]  K. Zsebo,et al.  Development of human mast cells from umbilical cord blood cells by recombinant human and murine c-kit ligand. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[47]  D. Metcalfe,et al.  Demonstration of the origin of human mast cells from CD34+ bone marrow progenitor cells. , 1991, Journal of immunology.

[48]  C. Kepley,et al.  Detection of MCT and MCTC types of human mast cells by immunohistochemistry using new monoclonal anti-tryptase and anti-chymase antibodies. , 1989, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[49]  L. Schwartz,et al.  Evaluation of human peripheral blood leukocytes for mast cell tryptase. , 1987, Journal of immunology.

[50]  L. Schwartz,et al.  Two types of human mast cells that have distinct neutral protease compositions. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[51]  M. Neuberger,et al.  A hapten-specific chimaeric IgE antibody with human physiological effector function , 1985, Nature.

[52]  G. Lazarus,et al.  Human skin chymotryptic proteinase. Isolation and relation to cathepsin g and rat mast cell proteinase I. , 1983, The Journal of biological chemistry.

[53]  S. Strobel,et al.  Human intestinal mucosal mast cells: evaluation of fixation and staining techniques. , 1981, Journal of clinical pathology.

[54]  T. Edgington,et al.  Lymphocyte collaboration is required for induction of murine monocyte procoagulant activity by immune complexes. , 1981, Journal of immunology.

[55]  K. Austen,et al.  Enzymes of the mast cell granule. , 1980, The Journal of investigative dermatology.

[56]  Y. Kitamura,et al.  Development of mast cells from grafted bone marrow cells in irradiated mice , 1977, Nature.

[57]  D. Conrad,et al.  Binding parameters of the interaction between rat IgE and rat mast cell receptors. , 1975, Journal of immunology.

[58]  L. Sachs,et al.  The induction of clones of normal mast cells by a substance from conditioned medium. , 1966, Experimental cell research.

[59]  L. Dahlin,et al.  Chitosan-film enhanced chitosan nerve guides for long-distance regeneration of peripheral nerves. , 2016, Biomaterials.

[60]  Joakim S. Dahlin,et al.  Mast cell progenitors: origin, development and migration to tissues. , 2015, Molecular immunology.

[61]  J. Arnhold,et al.  Differences in innate immune response between man and mouse. , 2014, Critical reviews in immunology.

[62]  Y. Okayama,et al.  Development, migration, and survival of mast cells , 2006, Immunologic research.

[63]  S. Bischoff,et al.  Isolation, culture, and characterization of intestinal mast cells. , 2006, Methods in molecular biology.

[64]  Robert E. Banfield,et al.  Techniques , 2005, Laboratory Investigation.

[65]  M. Eaman.  Immune system. , 2000, Nursing standard (Royal College of Nursing (Great Britain) : 1987).

[66]  D. Metcalfe,et al.  Mast cells cultured from the peripheral blood of normal donors and patients with mastocytosis originate from a CD34+/Fc epsilon RI- cell population. , 1994, Blood.

[67]  G. Nilsson,et al.  Phenotypic characterization of stem cell factor-dependent human foetal liver-derived mast cells. , 1993, Immunology.

[68]  G. Dewald,et al.  Establishment of an immature mast cell line from a patient with mast cell leukemia. , 1988, Leukemia research.