Off‑the‑shelf cryopreserved adipose tissue and ADSCs accelerate epithelial regeneration.

Delayed epithelialization remains a major limitation in wound repair. Adipose‑derived biomaterials offer regenerative advantages over other mesenchymal sources, but the epithelial healing potential of cryopreserved adipose tissue (AT) and adipose‑derived stem cells (ADSCs) remains unexplored, limiting their broader adoption as off‑the‑shelf therapies. This study evaluated the wound healing efficacy of post-thawed 2-month cryopreserved AT and mechanically isolated ADSCs (passaged 1) in in vivo rat and ex vivo human skin models. Wound closure was quantified by grid count, planimetry, histology, and 5-Chloromethylfluorescein diacetate (CMFDA) based live epithelial tracing. Cryo‑ADSC treatment achieved significantly faster early closure (65.31 ± 11.99% at day 3) than cryo‑AT (54.20 ± 20.40%) and controls (43.29 ± 10.99%) p < 0.05, with superior epithelial regeneration confirmed by planimetry and H&E till day 14 (p < 0.001). Ex vivo assays showed comparable trends, with cryo‑ADSCs promoting the greatest closure by day 7 (58.73 ± 20.55%, p < 0.001). CMFDA tracing revealed higher epithelial coverage than conventional histology, illustrating its value for dynamic assessment. These findings demonstrate that cryopreserved AT and ADSCs stored at -80 °C retain robust neo‑epithelial progression capacity, supporting their feasibility as scalable, off‑the‑shelf wound‑repair therapeutics.