
==== Front
Plast Reconstr Surg Glob Open
Plast Reconstr Surg Glob Open
GOX
Plastic and Reconstructive Surgery Global Open
2169-7574
Lippincott Williams & Wilkins Hagerstown, MD

GOX-D-24-00774
00048
10.1097/01.GOX.0001064000.77504.9a
3
PSTM Abstract Supplement
PSTM Top Abstracts/Posters 2024
Promoting Adipocyte Survival Using Embryonically Reset Adult Endothelial Cell Chaperones
Mayne Riley BS 1
Jeon Jini BA 1
Li Tim BA, BS 1
Salazar Martinez Hector MD 1
Dong Xue MD, PhD 1
Rafii Shahin 1
Spector Jason MD 1
Weill Cornell Medicine
9 2024
18 9 2024
12 Suppl 36-37Copyright © 2024 The Authors. Published by Wolters Kluwer Health, Inc. on behalf of The American Society of Plastic Surgeons. All rights reserved.
2024
https://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

OPEN-ACCESSTRUE
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pmcPURPOSES: Though widely used in plastic and reconstructive surgery autologous fat grafts are often limited by their unpredictable and often inadequate volume retention. A major cause of graft loss is the insufficient rate and degree of revascularization of the transplanted fat, resulting in ischemic necrosis. Previous work in our lab aimed at improving the survival of transplanted fat demonstrated that human umbilical vein endothelial cells (HUVEC) transduced with E4ORF1 region of human adenovirus Type 5 (E4s) promoted adipocyte survival in both in vitro and in vivo models. Here, we analyzed if a novel cell line, reset vascular cells (R-VEC), were capable of yielding similar results. This cell population consists of HUVEC adult endothelial cells that have been transduced with the transcription factor ETV2 which is responsible for promoting and maintaining microvascular network development. This transcription factor is normally only expressed during early angiogenesis and is no longer expressed after 20 weeks in utero. This unique cell line has demonstrated promising angiogenetic capabilities in in vitro culture, suggesting they might promote adipocyte survival after fat transplantation.

METHODS: Primary mature adipocytes were isolated from patient-derived fat tissue and cultured alone, with E4s, or with R-VECs within neutralized 0.6% (w/v) collagen. 200uL of each experimental group was plated in triplicate with an R-VEC density of 250,000 cells/mL. In adipocyte conditions, a concentration of 11.5% adipocyte (V/V) was mixed within collagen alone or with ECs in addition to collagen. Constructs were harvested on days 0, 4, and 7. Perilipin immunofluorescent staining around lipid droplets was used to verify live adipocytes. An entire cross-section of each construct was imaged and stitched using imaging software. Spaces between 70-120um (the average size of adipocytes), without perilipin expression were counted as non-viable adipocytes. The percentage of viable adipocytes was calculated.

RESULTS: Perilipin staining confirmed the presence of live adipocytes in all constructs and yielded consistent adipocyte survival rates at Day 0 across all groups. After 7 days, the R-VEC group exhibited significantly greater adipocyte viability when compared to both adipocytes in coculture with E4s and adipocytes in monoculture. On day 7, the R-VEC group demonstrated the highest adipocyte survival at 87.5%, with significantly lower amount in E4 at 63.4% and adipocyte alone at 49.5% (p<0.05).

CONCLUSIONS: In a biomimetic three-dimensional in vitro model of fat transplantation, co-culture of adipocytes with R-VECs resulted in superior adipocyte viability compared to culture with either E4 or HUVEC. These findings may have translational implications as R-VEC cells may be considered for future FDA authorization. Further studies are underway to determine if this pro-survival effect is resultant from the R-VEC capability to form microvascular networks, elaboration of pro-survival chemokines, or some combination of both.
