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In this work, we propose an optimized methodology to enhance the differentiation of 3T3-L1 fibroblasts into adipocytes. This process is crucial for adipocyte secretome release, which is fundamental for understanding the molecular mechanisms that are involved in obesity for in vitro studies. To achieve this, a pioglitazone dose-response assay was determined over a range varying from 0 to 10 \u00b5M. Lipid accumulation was evaluated using Oil-Red-O. The results showed that 10 \u00b5M pioglitazone enhanced differentiation and increased secretome production. This secretome was then added into two cell lines: PC3 and RAW264.7. In the PC3 cells, an increase of aggressiveness was observed in terms of viability and proliferation, with the increase of anti-inflammatory cytokines. Conversely, in RAW264.7 cells, a reduction of viability and proliferation was observed, with a decrease in the overexpression of pro-inflammatory cytokines. Overall, the present work constitutes an improved method for adipocyte secretome production that is suitable for experimental biology studies and that could help with our understanding of the molecular mechanisms underlying adiposity influence in other cells.<\/jats:p>","DOI":"10.3390\/biology11060806","type":"journal-article","created":{"date-parts":[[2022,5,24]],"date-time":"2022-05-24T22:04:06Z","timestamp":1653429846000},"page":"806","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Enhanced 3T3-L1 Differentiation into Adipocytes by Pioglitazone Pharmacological Activation of Peroxisome Proliferator Activated Receptor-Gamma (PPAR-\u03b3)"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6934-7641","authenticated-orcid":false,"given":"Catarina","family":"Teixeira","sequence":"first","affiliation":[{"name":"Laboratory of Medical and Industrial Biotechnology (LABMI), Porto 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Portugal"}]},{"given":"In\u00eas","family":"Santos","sequence":"additional","affiliation":[{"name":"Laboratory of Medical and Industrial Biotechnology (LABMI), Porto Research, Technology, and Innovation Center (PORTIC), Polytechnic Institute of Porto (IPP), 4200-374 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4127-1694","authenticated-orcid":false,"given":"Ana Catarina","family":"Rocha","sequence":"additional","affiliation":[{"name":"Laboratory of Medical and Industrial Biotechnology (LABMI), Porto Research, Technology, and Innovation Center (PORTIC), Polytechnic Institute of Porto (IPP), 4200-374 Porto, Portugal"},{"name":"Institute of Research, Innovation in Health (i3S), University of Porto, 4200-135 Porto, Portugal"},{"name":"Unit of Biochemistry (FMUP), Department of Biomedicine, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"}]},{"given":"In\u00eas","family":"Alencastre","sequence":"additional","affiliation":[{"name":"Institute of Research, Innovation in Health (i3S), University of Porto, 4200-135 Porto, Portugal"}]},{"given":"Ana Cl\u00e1udia","family":"Pereira","sequence":"additional","affiliation":[{"name":"Laboratory of Medical and Industrial Biotechnology (LABMI), Porto Research, Technology, and Innovation Center (PORTIC), Polytechnic Institute of Porto (IPP), 4200-374 Porto, Portugal"},{"name":"Institute of Research, Innovation in Health (i3S), University of Porto, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9709-9948","authenticated-orcid":false,"given":"Daniela","family":"Martins-Mendes","sequence":"additional","affiliation":[{"name":"Laboratory of Medical and Industrial Biotechnology (LABMI), Porto Research, Technology, and Innovation Center (PORTIC), Polytechnic Institute of Porto (IPP), 4200-374 Porto, Portugal"},{"name":"Institute of Research, Innovation in Health (i3S), University of Porto, 4200-135 Porto, Portugal"},{"name":"Unit of Biochemistry (FMUP), Department of Biomedicine, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Faculty of Health Sciences (FCS), Clinical Studies Center (CECLIN-HEFP), Fernando Pessoa Hospital, University Fernando Pessoa (UFP), 4420-096 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4537-5450","authenticated-orcid":false,"given":"Pedro","family":"Barata","sequence":"additional","affiliation":[{"name":"Laboratory of Medical and Industrial Biotechnology (LABMI), Porto Research, Technology, and Innovation Center (PORTIC), Polytechnic Institute of Porto (IPP), 4200-374 Porto, Portugal"},{"name":"Institute of Research, Innovation in Health (i3S), University of Porto, 4200-135 Porto, Portugal"},{"name":"Faculty of Health Sciences (FCS), Clinical Studies Center (CECLIN-HEFP), Fernando Pessoa Hospital, University Fernando Pessoa (UFP), 4420-096 Porto, 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