May 15, 2026
TDA-MIT: Establishment of the mouse wound healing model and sample collection
- Ke Zhang1,
- Chia-Kang Ho2
- 1Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA;
- 2Krantz Family Center for Cancer Research, Massachusetts General Hospital, Boston, MA, USA
- Cellular Senescence Network (SenNet) Method Development Community
Protocol Citation: Ke Zhang, Chia-Kang Ho 2026. TDA-MIT: Establishment of the mouse wound healing model and sample collection. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvoeewbl4o/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 14, 2026
Last Modified: May 15, 2026
Protocol Integer ID: 317164
Keywords: establishment of the mouse wound healing model, mouse wound healing model, thickness excisional wound healing model, skin wound tissue, cryopreservation procedure, frozen on dry ice, tissue collection, dry ice, including tissue collection, early healing
Abstract
This protocol describes the establishment of a mouse full-thickness excisional wound healing model in young and aged p16-3MR mice, including tissue collection and cryopreservation procedures. Skin wound tissues were collected at baseline and during early healing, embedded in OCT, snap-frozen on dry ice, and stored at −80 °C for downstream spatial and molecular analyses.
Safety warnings
This protocol requires prior approval by the users' Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee.
Wound healing mouse model
Full-thickness excisional wounds were generated on the dorsal skin of 2-month-old and 24-month-old p16-3MR mice. Prior to surgery, mice were anesthetized with 3% isoflurane via nose cone and administered subcutaneous analgesia using Ethiqa XR (3.25 mg/kg, 1.3 mg/mL) 30 minutes before wounding. The dorsal area was first shaved with electric clippers and then depilated using a chemical depilatory cream, which was applied for no more than 30 seconds and thoroughly removed with moist gauze to avoid skin irritation. A single 6-mm full-thickness wound, including panniculus carnosus, was created on the central dorsum using a sterile biopsy punch. For each mouse, two samples were collected: (1) the excised wound tissue at day 0, and (2) wound-edge skin at day 3 post-injury using a 6-mm punch biopsy centered at the healing margin. All procedures were conducted in compliance with institutional animal care and use guidelines.
Dissected mouse skin samples were placed into OCT-containing plastic molds, frozen on dry ice, and stored at -80°C.
Frozen OCT-embedded tissue blocks were cryosectioned for downstream analyses. Tissue sections were used for spatial sequencing with Raman imaging.