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In this work, we validated a dynamic 2D breast-on-a-chip microfluidic bioassay operated at a controlled infusion rate of 20 \u00b5L\/h to assess anticancer drug responses under defined flow conditions. Using Palbociclib as a reference compound, we evaluated proliferation, viability\/apoptosis, cytoskeleton organization, and differential processing of the resistance-associated marker PARP1 in MCF-7 and T47D breast cancer cells. Under dynamic microfluidic conditions, Palbociclib induced dose-dependent effects, with the higher concentration (20 \u00b5M) consistently reducing cell proliferation and viability and increasing late apoptosis compared to 10 \u00b5M Palbociclib. Cytoskeletal disorganization was observed at both concentrations, while differential PARP1 processing patterns between MCF-7 and T47D cells were detected across doses. These responses are consistent with known effects of CDK4\/6 inhibition and were reproducibly captured under controlled flow conditions. Overall, our results demonstrate that this breast-on-a-chip microfluidic model provides a reproducible and physiologically relevant in vitro platform for integrated assessment of drug efficacy and resistance-associated markers under dynamic perfusion.<\/jats:p>","DOI":"10.3390\/cells15050446","type":"journal-article","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T16:06:59Z","timestamp":1772467619000},"page":"446","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Breast-on-a-Chip Microfluidic Model to Assess the Effect of Palbociclib in MCF-7 and T47D Cancer Cells"],"prefix":"10.3390","volume":"15","author":[{"given":"Ingrid Larissa Melo","family":"Souza","sequence":"first","affiliation":[{"name":"Laboratory for Applied Science and Technology in Health (LACTAS), Carlos Chagas Institute, Fiocruz, Curitiba 81350-010, PR, 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Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sthefanie Ribas","family":"Klein","sequence":"additional","affiliation":[{"name":"Laboratory for Applied Science and Technology in Health (LACTAS), Carlos Chagas Institute, Fiocruz, Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ma\u00edra Barbosa","family":"e Reis","sequence":"additional","affiliation":[{"name":"Laboratory for Applied Science and Technology in Health (LACTAS), Carlos Chagas Institute, Fiocruz, Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andr\u00e9ia Akemi","family":"Suzukawa","sequence":"additional","affiliation":[{"name":"Laboratory of Basic Biology of Stem Cells (Labcet), Carlos Chagas Institute, Fiocruz, Curitiba 81350-010, PR, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9397-4127","authenticated-orcid":false,"given":"Dalila 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