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Staged Dermal-Hypodermal Xenografting to Model Trilaminar Human Skin in the Murine Host

Shawn Loder, MD; Phoebe Lee, BS; Patricia Leftwich; MS; Wayne Nerone, BA; Kacey Marra, PhD; Lauren Kokai, PhD, and J. Peter Rubin, MD
University of Pittsburgh
2021-02-01

Presenter: Shawn Loder

Affidavit:
Certified

Director Name: Vu T. Nguyen, MD

Author Category: Resident Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction

Introduction:
Reliability and relevance of our pre-clinical models represents one of the most persistent needs in plastic surgery. Unfortunately, the inherent complexity and unique architecture of human skin directly limits the applicability of lower animals to human practice. Human skin xenografts present one potential solution to; however, traditional skin grafting lacks the stem-cell rich hypodermis. Here we propose a strategy utilizing staged engraftment to recreate trilaminar human skin in a murine host.

Methods:
Full-thickness skin and lipoaspirate was collected from panniculectomy samples. At time of harvest lipoaspirate samples were grafted to the flank of athymic mice. We banked 1 cm2 full-thickness skin for 7 days in McCoy's Media. After 7 days banked skin was grafted overlying the established xenograft fat pad. At 1-6 weeks animals were sacrificed. Grafts were collected for histologic and viability analysis.

Results:
Viability of cultured skin was maintained at 83.6+/-0.95% at 7 days. Xenograft fat pads demonstrated gross formation of a de novo vascular network by 1-week post-engraftment. Xenografted skin demonstrated viability of 90.5+/-1.6% at 6 weeks post-engraftment. Gross histology demonstrated re-integration of adipose and dermal layers.

Conclusion:
In vivo modelling of trilaminar skin is a critical need in plastic surgery. Several methods including use of porcine and/or tissue engineered systems have been proposed and/or used each with their own risks and limitations. Here we describe a simple two-step staged engraftment model supported by short term tissue banking to generate donor-matched xenograft with a robust and viable hypodermal layer consistent with native human skin architecture.

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