Plastic Surgery Research Council
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EX-VIVO ASSEMBLY AND TRANSPLANTATION OF A STEM CELL-DERIVED HYBRID GRAFT USING THE LGR6+ EPITHELIAL STEM CELL ON COMMON ACELLULAR MATRICES FOR TREATMENT OF FULL THICKNESS SOFT TISSUE WOUNDS
Presenter: Denver M Lough, MD, PhD
Co-Authors: Cooney DS; Mendenhall SD; Yang M; Reichensperger JD; Cox LA; Cosenza NM; Wetter N; Harrison CE; Neumeister MW
SIU School of Medicine

Background: Prior research shows that the migration of Leucine-rich repeat-containing G-protein coupled Receptor epithelial stem cells (LGR5+ and LGR6+) into wounds augments healing and permits nascent hair growth in areas devoid of these cells following local full thickness soft tissue trauma. Here, we assess the LGR-expressing cell s potential as a hybrid graft in full thickness soft tissue wounds with the goal of developing an easily assembled biological dressing to provide patients with their own focal stem cell (SC) population.

Methods: GFP expressing LGR6+ epithelial stem cells (SC) were isolated from C57BL/6(UBC-GFP) mice using FACS gated on CD34, CD73, and LGR6. These cells were then seeded on a variety of acellular matrices. Viability and attachment was assessed using confocal and scanning electron microscopy and bioluminescence. LRG6+ SC seeded matrices were placed into acute 3rd degree burn wound beds of Nu/Nu mice and monitored for viability, wound healing, and growth factor expression. Bioluminescent imaging supported SC confluence within the graft as well as transplant viability. Healing rates were documented and compared between mice receiving the SC enriched grafts vs. control. On days 0, 5, 10 and 15, grafted areas were harvested and assessed for gross, microscopic and molecular healing using digital photography, immunofluorescence, RT-PCR, and angiogenic protein production.

Results: LGR6+ seeded matrices show significantly enhanced gross wound healing and upregulation of key WNT, EGF, VEGF and angiogenesis mRNA transcripts and peptides, when compared to matrix-only and negative controls.

Conclusion: Here, we suggest a novel role for an LGR6+ epithelial stem cell enriched hybrid graft for the development of a fully transplantable living matrix for use in tissue engineering and reconstructive transplantation. Furthermore, with epithelium being a major source of alloantigenicity, this system can be used to develop a focus of chimeric epithelial expansion for engraftment onto more complex composite tissue allografts.


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