Mouse experiments were performed according to Good Scientific Practice-principles and approved by the Ethical Committee (EC) of the Faculty of Veterinary Medicine, Ghent University (EC 2020-030 and 2021-018).
Eight-week (w)-old female and male BALB/c mice were mated and pups were weaned 12-14 days (d) post-parturition. One hour (h) after weaning, lactating females were intraductally inoculated in the third mammary gland pair with 5 × 104 4T1-luc cells suspended in a 100 μl mixture of PBS and Matrigel® (1:10; Corning, Bedford, MA, USA) under inhalation anesthesia and analgesia as described previously (15 (link)). For intravenous (i.v.) treatment, eMSCs or eEpSCs were dissolved in DMEM at 3x105 per 100 μl and injected through the tail vein using a 29G insulin needle. I.v. injections with DMEM only were used as a negative (sham) control. Antibody treatments for depletion of CD4+ (clone GK1.5) and CD8α+ T-cells (clone YTS169.4) or rat IgG2b isotype controls (clone LTF-2) were purchased from BioXCell (West Lebanon, NH, USA) and diluted (200 μg/100 μl) in InVivoPure dilution buffer (BioXCell) at pH6.5 (for anti-CD4) or pH7 (for anti-CD8α and isotype control) for i.p. administration.
Eight-week (w)-old female and male BALB/c mice were mated and pups were weaned 12-14 days (d) post-parturition. One hour (h) after weaning, lactating females were intraductally inoculated in the third mammary gland pair with 5 × 104 4T1-luc cells suspended in a 100 μl mixture of PBS and Matrigel® (1:10; Corning, Bedford, MA, USA) under inhalation anesthesia and analgesia as described previously (15 (link)). For intravenous (i.v.) treatment, eMSCs or eEpSCs were dissolved in DMEM at 3x105 per 100 μl and injected through the tail vein using a 29G insulin needle. I.v. injections with DMEM only were used as a negative (sham) control. Antibody treatments for depletion of CD4+ (clone GK1.5) and CD8α+ T-cells (clone YTS169.4) or rat IgG2b isotype controls (clone LTF-2) were purchased from BioXCell (West Lebanon, NH, USA) and diluted (200 μg/100 μl) in InVivoPure dilution buffer (BioXCell) at pH6.5 (for anti-CD4) or pH7 (for anti-CD8α and isotype control) for i.p. administration.
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