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An Alternative Method for Harvest and Processing Fat Grafts: An In Vitro Study of Cell Viability and Survival

Gonzalez, Andrea Moreira M.D.; Lobocki, Catherine M.S.; Kelly, Christopher P. M.D.; Jackson, Ian T. M.D., D.Sc.(Hon.)

Plastic & Reconstructive Surgery:
doi: 10.1097/01.prs.0000264401.19469.ad
Cosmetic: Original Articles
Abstract

Background: Adipocyte viability has been emphasized as essential for fat graft survival. There is no universal agreement on the methodology for handling fat grafts.

Objective: Two different methods of fat harvesting and techniques of tissue processing were compared by assessing cell viability, damage, and growth in vitro.

Method: Fat was harvested from Zucker rats (n = 5) using (1) a 3-mm liposuction cannula with a 60-cc syringe (right side) or (2) a 2-mm blunt needle with a 10-cc syringe adapted to a fine-needle aspiration apparatus (left side). Tissues were then processed by decantation or cotton towel drying. Five samples for each of the four techniques were studied. Fat samples were processed for culture, and adipocytes and preadipocytes were plated in culture medium and expanded in vitro. Cell viability was assessed using cell counts, the MTT proliferation assay, G3PDH activity, and Oil Red O stain.

Results: Method 1 exerted significantly higher pressure (p = 0.009) than method 2 (643 ± 2.5 versus 537 ± 13.6 mm Hg). A larger oil layer was apparent with method 1 (1.11 ± 0.29 g) than with method 2 (0.56 ± 0.28 g). In addition, the highest number of viable preadipocytes was obtained using method 2B (p = 0.017). In culture, preadipocytes plated in 4F differentiation medium started to differentiate after 1 week, while those in Dulbecco's modified Eagle's medium/F12 with serum proliferated but did not differentiate. Mature adipocytes in adipogenic medium dedifferentiated and later redifferentiated into fat cells.

Conclusions: Fat viability was better when fat was harvested by fine-needle aspiration. The plasticity of mature adipocytes and preadipocytes in vitro suggested that both might be involved in fat graft integration.

Author Information

Cleveland, Ohio; Southfield, Mich.; and Seattle, Wash.

From the Department of Plastic Surgery, Cleveland Clinic Foundation; Research Department and Institute for Craniofacial and Reconstructive Surgery, Providence Hospital; and Virginia Mason Medical Center.

Received for publication November 15, 2005; accepted March 23, 2006.

Presented at the 82nd Annual Meeting of the American Association of Plastic Surgeons, in Baltimore, Maryland, May 4 through 7, 2003.

Ian T. Jackson, M.D., Craniofacial and Reconstructive Institute, 16001 West Nine Mile Road, 3rd Floor, Fisher Center, Southfield, Mich. 48075, ianjackson@juno.com

©2007American Society of Plastic Surgeons