Op site flexigridtm

Ly5.1 mice were anaesthetised with Ketamine (100 mg/kg) and Xylazil (10 mg/kg) (Troy Laboratories). The flanks of the mice were shaved using clippers and depilated with Veet (Reckitt Benckiser), and the skin (~2 mm²) was abraded using a MultiPro Dremel with a grindstone attachment. B16.F10‐OVA‐expressing melanoma cells (1 × 10⁵) were resuspended in 10 μl of Matrigel™ (BD Biosciences) and orthotopically applied to the lesion. Set Matrigel was covered with a piece of Op‐site FlexigridTM (Smith and Nephew). The torsos of the mice were wrapped with a soft hypoallergenic MicroporeTM tape (3M Health Care) and then by a stronger porous polyethylene TransporeTM tape (3M Health Care) to protect the abraded site. Twenty‐four hours later, 2 × 10⁵ FACS‐purified CD8⁺ naive CD44^loCD62L^hi OT‐1 T cells were adoptively transferred. Recipient mice were monitored daily until bandages were removed 6 days after tumour engraftment.

Mice were anaesthetized with a mixture of ketamine (100 mg/kg) and xylazil (10 mg/kg) (Troy Laboratories) administered intraperitoneally (10 mL/g body weight).
Refresh®Lacri-Lube® (Allergan) was applied to mouse eyes for the duration of anaesthesia. The flank of the mouse was shaved and depilated with Veet®(Reckitt Benckiser). A small (~2 mm²) area of surface skin on the flank was abraded using a MultiPro Dremel with a grindstone attachment on low speed. B16 melanoma cells (10⁵) were washed, resuspended in 10 μL of Matrigel™ (BD), applied to the abraded area and allowed to set. To contain the cells, the abraded site was covered with a piece of Op-site FlexigridTM (Smith and Nephew), then the torsos of the mice were wrapped, first with a soft hypoallergenic MicroporeTM tape (3 M Health Care) and then a second layer of Fixomull Stretch (BSN Healthcare). Mice recovered from anaesthesia on a heating pad and were monitored daily until bandages were removed 5 days post-grafting. Mice with tumours >1000 mm³ were euthanised. Tumour-free mice were defined as mice with no palpable masses.