{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T04:27:21Z","timestamp":1787113641044,"version":"build-2736575974"},"reference-count":31,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T00:00:00Z","timestamp":1786665600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001823","name":"Ministry of Education Youth and Sports","doi-asserted-by":"publisher","award":["CZ.02.01.01\/00\/23_020\/0008499"],"award-info":[{"award-number":["CZ.02.01.01\/00\/23_020\/0008499"]}],"id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]},{"award":["CZ.02.01.01\/00\/23_020\/0008499"],"award-info":[{"award-number":["CZ.02.01.01\/00\/23_020\/0008499"]}],"id":[{"id":"https:\/\/ror.org\/037n8p820","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Therapies based on mesenchymal stromal cells (MSCs) have high potential in the field of regenerative medicine due mainly to their immunomodulatory properties. However, their clinical translation is hampered by a lack of sufficiently standardised potency tests. Since macrophages constitute key mediators of the effects of MSCs, macrophage-based assays potentially provide a relevant in vitro tool for the evaluation of the activity of MSC products. This study involved the coculturing of canine adipose-derived mesenchymal stromal cells (ASCs) with macrophages derived from human THP-1 and U937 monocyte cell lines, murine RAW264.7 macrophages and primary human macrophages. The M2 polarisation was assessed following stimulation with IL-4\/IL-13 in THP-1 and U937 macrophages. The mRNA expression of the pro- and anti-inflammatory markers was analysed using qPCR. The ASC transwell coculture altered the LPS-induced inflammatory mRNA expression in a strongly model- and marker-dependent manner. The U937-derived macrophages exhibited the most consistent suppression of the tested inflammatory transcripts and the RAW264.7 cells provided a practical readout for selected inflammatory markers, whereas the THP-1 macrophages evinced the suppression of TNFA but not IL1B or PTGS2 under the selected stimulation conditions. IL-4\/IL-13 induced moderate but statistically non-significant changes in IL10 and TGFB1 in the U937-derived macrophages but no reproducible response in the THP-1-derived macrophages. In a subsequent U937 coculture experiment, ASC-derived paracrine factors altered selected M2-associated transcripts at specific time points. The results thus provided support for macrophage-based transcriptional readouts as an early-stage tool for comparing responder macrophage models and detecting the selected anti-inflammatory paracrine effects of canine ASCs; the U937 cells were found to be particularly suitable for the study of inflammatory polarisation and the RAW264.7 cells for the purpose of standardised screening.<\/jats:p>","DOI":"10.3390\/biom16081190","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T15:48:36Z","timestamp":1786722516000},"page":"1190","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Macrophage-Based Transcriptional Assays for the Comparative Assessment of the Anti-Inflammatory Paracrine Activity of Canine Adipose-Derived Mesenchymal Stromal Cells"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-7249-5201","authenticated-orcid":false,"given":"Andrea","family":"Exnerov\u00e1","sequence":"first","affiliation":[{"name":"Cell Physiology Group, Contipro a.s., Doln\u00ed Dobrou\u010d 401, 562 01 Doln\u00ed Dobrou\u010d, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabina","family":"Seidlov\u00e1","sequence":"additional","affiliation":[{"name":"Cell Physiology Group, Contipro a.s., Doln\u00ed Dobrou\u010d 401, 562 01 Doln\u00ed Dobrou\u010d, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-4205-060X","authenticated-orcid":false,"given":"V\u011bra","family":"Da\u0148kov\u00e1","sequence":"additional","affiliation":[{"name":"Cell Physiology Group, Contipro a.s., Doln\u00ed Dobrou\u010d 401, 562 01 Doln\u00ed Dobrou\u010d, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6575-9726","authenticated-orcid":false,"given":"Vojt\u011bch","family":"Pavl\u00edk","sequence":"additional","affiliation":[{"name":"Cell Physiology Group, Contipro a.s., Doln\u00ed Dobrou\u010d 401, 562 01 Doln\u00ed Dobrou\u010d, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7864-7689","authenticated-orcid":false,"given":"Kristina","family":"Ne\u0161porov\u00e1","sequence":"additional","affiliation":[{"name":"Cell Physiology Group, Contipro a.s., Doln\u00ed Dobrou\u010d 401, 562 01 Doln\u00ed Dobrou\u010d, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1186\/s13287-022-03054-0","article-title":"Clinical Application of Mesenchymal Stem Cell in Regenerative Medicine: A Narrative Review","volume":"13","author":"Margiana","year":"2022","journal-title":"Stem Cell Res. 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