Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators
Introduction: Mast cells are highly granulated tissue-resident leukocytes that require a three-dimensional matrix to differentiate and mediate immune responses. However, almost all cultured mast cells rely on two-dimensional suspension or adherent cell culture systems, which do not adequately reflec...
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| Format: | Article |
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Frontiers Media S.A.
2023-04-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1160460/full |
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| author | Marianna Kulka Marianna Kulka Ashley Wagner Jae-Young Cho Syed Benazir Alam Joy Ramielle Santos Juan Jovel Leshern Karamchand Marcelo Marcet-Palacios |
| author_facet | Marianna Kulka Marianna Kulka Ashley Wagner Jae-Young Cho Syed Benazir Alam Joy Ramielle Santos Juan Jovel Leshern Karamchand Marcelo Marcet-Palacios |
| author_sort | Marianna Kulka |
| collection | DOAJ |
| description | Introduction: Mast cells are highly granulated tissue-resident leukocytes that require a three-dimensional matrix to differentiate and mediate immune responses. However, almost all cultured mast cells rely on two-dimensional suspension or adherent cell culture systems, which do not adequately reflect the complex structure that these cells require for optimal function.Methods: Crystalline nanocellulose (CNC), consisting of rod-like crystals 4–15 nm in diameter and 0.2–1 µm in length, were dispersed in an agarose matrix (12.5% w/v), and bone marrow derived mouse mast cells (BMMC) were cultured on the agarose/CNC composite. BMMC were activated with the calcium ionophore A23187 or immunoglobulin E (IgE) and antigen (Ag) to crosslink high affinity IgE receptors (FcεRI).Results: BMMC cultured on a CNC/agarose matrix remained viable and metabolically active as measured by reduction of sodium 3′-[1-[(phenylamino)-carbony]-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate (XTT), and the cells maintained their membrane integrity as analyzed by measuring the release of lactate dehydrogenase (LDH) and propidium iodide exclusion by flow cytometry. Culture on CNC/agarose matrix had no effect on BMMC degranulation in response to IgE/Ag or A23187. However, culture of BMMC on a CNC/agarose matrix inhibited A23187-and IgE/Ag-activated production of tumor necrosis factor (TNF) and other mediators such as IL-1β, IL-4, IL-6, IL-13, MCP-1/CCL2, MMP-9 and RANTES by as much as 95%. RNAseq analysis indicated that BMMC expressed a unique and balanced transcriptome when cultured on CNC/agarose.Discussion: These data demonstrate that culture of BMMCs on a CNC/agarose matrix promotes cell integrity, maintains expression of surface biomarkers such as FcεRI and KIT and preserves the ability of BMMC to release pre-stored mediators in response to IgE/Ag and A23187. However, culture of BMMC on CNC/agarose matrix inhibits BMMC production of de novo synthesized mediators, suggesting that CNC may be altering specific phenotypic characteristics of these cells that are associated with late phase inflammatory responses. |
| first_indexed | 2024-04-09T18:39:56Z |
| format | Article |
| id | doaj.art-a7114b8d42e24d85b073e3885dadb70d |
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| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-09T18:39:56Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-a7114b8d42e24d85b073e3885dadb70d2023-04-11T06:02:31ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-04-011110.3389/fbioe.2023.11604601160460Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediatorsMarianna Kulka0Marianna Kulka1Ashley Wagner2Jae-Young Cho3Syed Benazir Alam4Joy Ramielle Santos5Juan Jovel6Leshern Karamchand7Marcelo Marcet-Palacios8Nanotechnology Research Centre, National Research Council Canada, Edmonton, AB, CanadaDepartment of Medical Microbiology and Immunology 6-020 Katz Group Centre, University of Alberta, Edmonton, AB, CanadaNanotechnology Research Centre, National Research Council Canada, Edmonton, AB, CanadaNanotechnology Research Centre, National Research Council Canada, Edmonton, AB, CanadaNanotechnology Research Centre, National Research Council Canada, Edmonton, AB, CanadaDepartment of Medicine, University of Alberta, Edmonton, AB, CanadaThe Metabolomics Innovation Centre (TMIC), 7-12 Heritage Medical Research Centre, University of Alberta, Edmonton, AB, CanadaNanotechnology Research Centre, National Research Council Canada, Edmonton, AB, CanadaNorthern Alberta Institute of Technology, Edmonton, AB, CanadaIntroduction: Mast cells are highly granulated tissue-resident leukocytes that require a three-dimensional matrix to differentiate and mediate immune responses. However, almost all cultured mast cells rely on two-dimensional suspension or adherent cell culture systems, which do not adequately reflect the complex structure that these cells require for optimal function.Methods: Crystalline nanocellulose (CNC), consisting of rod-like crystals 4–15 nm in diameter and 0.2–1 µm in length, were dispersed in an agarose matrix (12.5% w/v), and bone marrow derived mouse mast cells (BMMC) were cultured on the agarose/CNC composite. BMMC were activated with the calcium ionophore A23187 or immunoglobulin E (IgE) and antigen (Ag) to crosslink high affinity IgE receptors (FcεRI).Results: BMMC cultured on a CNC/agarose matrix remained viable and metabolically active as measured by reduction of sodium 3′-[1-[(phenylamino)-carbony]-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate (XTT), and the cells maintained their membrane integrity as analyzed by measuring the release of lactate dehydrogenase (LDH) and propidium iodide exclusion by flow cytometry. Culture on CNC/agarose matrix had no effect on BMMC degranulation in response to IgE/Ag or A23187. However, culture of BMMC on a CNC/agarose matrix inhibited A23187-and IgE/Ag-activated production of tumor necrosis factor (TNF) and other mediators such as IL-1β, IL-4, IL-6, IL-13, MCP-1/CCL2, MMP-9 and RANTES by as much as 95%. RNAseq analysis indicated that BMMC expressed a unique and balanced transcriptome when cultured on CNC/agarose.Discussion: These data demonstrate that culture of BMMCs on a CNC/agarose matrix promotes cell integrity, maintains expression of surface biomarkers such as FcεRI and KIT and preserves the ability of BMMC to release pre-stored mediators in response to IgE/Ag and A23187. However, culture of BMMC on CNC/agarose matrix inhibits BMMC production of de novo synthesized mediators, suggesting that CNC may be altering specific phenotypic characteristics of these cells that are associated with late phase inflammatory responses.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1160460/fullmast cellnanocellulosecytokinesinflammationIgE |
| spellingShingle | Marianna Kulka Marianna Kulka Ashley Wagner Jae-Young Cho Syed Benazir Alam Joy Ramielle Santos Juan Jovel Leshern Karamchand Marcelo Marcet-Palacios Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators Frontiers in Bioengineering and Biotechnology mast cell nanocellulose cytokines inflammation IgE |
| title | Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators |
| title_full | Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators |
| title_fullStr | Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators |
| title_full_unstemmed | Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators |
| title_short | Agarose/crystalline nanocellulose (CNC) composites promote bone marrow-derived mast cell integrity, degranulation and receptor expression but inhibit production of de novo synthesized mediators |
| title_sort | agarose crystalline nanocellulose cnc composites promote bone marrow derived mast cell integrity degranulation and receptor expression but inhibit production of de novo synthesized mediators |
| topic | mast cell nanocellulose cytokines inflammation IgE |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1160460/full |
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