MouseWoundC57BL/6J plus fibroblast lineage reporter and conditional lines including En1-Cre and Wnt1-Cre crossed to dual fluorescence membrane and nuclear reporters, N-cadherin (Cdh2) floxed mice and Cas9 knock-in miceBoth male and female mice; human donors of both sexesAddition to explant culture medium for Exherin; subcutaneous injection into the fascia surrounding wounds for AAV6 vectorsExherin 500 microgram/mL in explant culture medium; AAV6 vi…Exherin present in the medium from the start of explant culture, that is from t…
Model: Full-thickness excisional skin wounds through the panniculus carnosus and fascia (2 mm, 3 mm and 5 mm biopsy punches, some splinted with silicone rings), intravital imaging through a dorsal skinfold chamber, and ex vivo full-thickness skin-fascia explants (scar-like tissue in a dish, SCAD) from back skin, buccal mucosa and human skin
Injury triggered N-cadherin upregulation and swarming collective migration of Engrailed-1 lineage fascia fibroblasts towards the wound centre, a behaviour absent from oral mucosa fibroblasts. Exherin prevented fibroblast aggregation and skin contraction, abolished wound-induced alpha-SMA upregulati…
