Female c57bl 6 mice

Female C57BL/6 mice were obtained at 6 weeks of age from Janvier Labs (France) and housed at the Department of Experimental Medicine, University of Copenhagen. All procedure were approved by the National Animal Experiments Inspectorate. E.G7-OVA and B16-OVA tumors were established in C57BL/6 mice by s.c. injection of 3 × 10⁵ viable cells (day 0). Tumors were consequently allowed to establish for 7 days for E.G7-OVA and 10 days for B16-OVA before initiation of treatment. Before inclusion, mice were randomized according to tumor volume.

Female C57BL/6 mice were obtained from Janvier Labs (Paris, France). NMRI mice were obtained either from Janvier or Charles River Labs (Sulzfeld, Germany). All mice were maintained under specific pathogen-free conditions. At the start of individual experiments mice were age-matched and 6–8 weeks of age.
All experiments were carried out with the rodent parasite Plasmodium berghei ANKA (PbANKA). PbANKA sporozoites were isolated by dissection of salivary glands (SGs) from female Anopheles stephensi mosquitoes at days 17–21 after a bloodmeal on infected NMRI mice. To obtain Pb ANKA radiation-attenuated sporozoites (PbRAS), isolated sporozoites were treated by exposure to 150 Gy of X-rays (X-rad 320, Precision X-Ray, University Hospital Heidelberg).

Female C57Bl/6 mice, 6–7 weeks old and weighing 20-25 g were obtained from Janvier Labs (Le Genest St. Isle, France). C57Bl/6-CD11c-GFP-DTR, C57Bl/6-Lang-GFP mice (a generous gift of Dr. B. Malissen, CIML, Marseille, France) and TLR9^−/− mice (gift from Invivogen, Toulouse, France) were bred in our animal facility. Nude mice were purchased from Charles River Laboratories France (L’Arbresle, France). Hspa1b-LucF (+/+) Hspa1b-mPlum (+/+) mice were obtained from F. Couillaud (EA7435, Université de Bordeaux, France). Hspa1b-LucF (+/+) Hspa1b-mPlum (+/+) mice contained a transgene that allowed firefly luciferase and mPlum fluorescent protein expression under the control of thermo-inducible heat-shock protein (Hsp70) promoter 1B (Hspa1b) [36 (link)]. All experiments involving animals were approved by the ethic committee (n°APAFIS#4268-2016022516345743v2) and performed in compliance with relevant laws and institutional guidelines.

Female C57BL/6 mice aged 8–12 weeks were purchased from Janvier Labs (Saint Berthevin, France). All mice were maintained on a standard diet with food and water ad libitum and a 12-h light-day-night rhythm. Under isoflurane anesthesia (0.6 mL/L oxygen and 2.5% isoflurane), 4 mL sterile air was injected into the shaved skin at the level of the scapulae of each mouse to create a dorsal subcutaneous air-pouch. The air-pouches were inflated with 2 mL of sterile air on days 2 and 4. On the fifth day, the biomaterial was implanted in a minimally invasive procedure under isoflurane anesthesia using an in-house, easy-to-use application system (Figures 1A, B). This consisted of a 5 mL syringe (Discardit; Becton Dickinson, United States) and a 21 ½-G cannula (MicrolanceTM 3; Becton Dickinson, United States). The implant was inserted into the cannula under sterile conditions before being inserted into the air-pouch. The implants were left in the mice for 6 h to analyze the short-term response or for 10 days to analyze the long-term response (short-term: n = 6 animals per experimental group and time, long-term: n = 5 animals per control group and time). Animals receiving phosphate buffered saline (PBS) served as controls.

Female C57BL6 mice were obtained from Janvier. EpSCs were isolated from telogen back skin collected from P56-60 mice, as previously described [27 (link)] with the following changes: fat and muscle tissue were removed from back skin using a scalpel. The skin was incubated in 0.5% Trypsin-EDTA (10X; Gibco,15400054) for 25 min at 37 °C on an orbital shaker. A single-cell suspension was then obtained by scraping the skin with a scalpel followed by neutralizing the trypsin by adding 1X PBS buffer containing 2% chelexed FBS (PBS-FBS(-)) (Gibco; 10010-015). The resulting cell suspension was then filtered through 70 µm and 40 μm cell strainers (Corning; 431750, 431751) and spun down. EpSCs were isolated using magnetic-associated cell sorting using Anti-SCA-1 microbeads (Miltenyi Biotec, 130-106-641) and a MACS MultiStand system (Miltenyi Biotec) together with MS columns (Miltenyi Biotec, 130-042-201). The resulting SCA-1+ EpSCs were spun down and resuspended in Trizol LS (Thermo Fisher, 102960-10). RNA was isolated using the Direct-zol™ RNA MiniPrep kit (Zymo Research, R2050) and concentration was determined using the Qubit^TM RNA BR assay kit (Invitrogen, Q10210). All animal procedures were approved by the Veterinary Office of the Canton of Zurich, Switzerland (License ZH233/2019).