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Promoting Adipocyte Survival In An In Vitro Model Of Fat Grafting Using Embryonically Reset Adult Endothelial Cell Chaperones
Abby Chopoorian Fuchsman
*, Sophia Salingaros, Kate Manley, Riley Mayne, Matthew W. Liao, Samuel Medina, Xue Dong, Jason A. Spector
Weill Cornell Medicine, New York, NY
Background: Fat grafting is limited by inconsistency in volume retention. We have previously shown that co-culturing adipocytes with reset vascular endothelial cells (R-VECs), which are human umbilical vein endothelial cells (HUVECs) transduced with the angiogenic transcription factor ETV2, promotes adipocyte survival in vitro. The present study aims to compare adipocyte retention of adipocyte monoculture to co-cultures with R-VECs and wildtype HUVECs at "high" and "low" concentrations to investigate the impact of cell density on adipocyte survival.
Methods: Primary adipocytes extracted from donor adipose tissue or lipoaspirate were cultured alone, with HUVECs at "low" (250,000 cell/mL) and "high" (2.5 million cells/mL) concentrations, or with R-VECs at the same concentrations. Co-cultures were plated by mixing a solution of 13% adipocytes (V/V) with either 0.6% neutralized type I plain collagen alone, or with collagen and supplemented with R-VECs or HUVECs. Constructs were fixed after 0, 4, and 7 days. Circular signals within the proper size range and morphology for adipocytes were identified as either viable (perilipin expression) or non-viable (no perilipin expression) adipocytes/lipid droplet. Percentage of live adipocytes was calculated.
Results: Low-density and high-density R-VEC co-cultures exhibited higher percent adipocyte viability compared to monoculture at days 4 (47.7 vs 31.6 [p<0.0001] and 49.9 vs 31.6 [p=0.0124]) and 7 (39.3 vs 24.0 [p=0.0006] and 39.6 vs 24.0 [p=0.0125]). Low-density HUVEC co-culture yielded higher viability compared to monoculture at days 4 (53.3 vs 31.6 [p<0.0001]) and 7 (51.8 vs 24.0 [p<0.0001]).
Conclusions: Co-culturing adipocytes with endothelial cell chaperones significantly improved adipocyte viability in vitro, showing promise as a potential adjunct to improve fat grafting. There were no differences between R-VEC and HUVEC groups, suggesting that the effect of endothelial cell chaperones on adipocyte survival is not directly proportional to increased angiogenic capability of the R-VECS.
Adipocyte viability of Monoculture (AO), Low-Density R-VEC (L2) and HUVEC (LH), and High-Density R-VEC (H2) and HUVEC (HH) cocultures at Days 0, 4, and 7.
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