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In Vitro Evaluation of Different Methods of Handling Human Liposuction Aspirate and Their Effect on Adipocytes and Adipose Derived Stem Cells
Author(s) -
Palumbo Paola,
Miconi Gianfranca,
Cinque Benedetta,
la Torre Cristina,
Lombardi Francesca,
Zoccali Giovanni,
Orsini Gino,
Leocata Pietro,
Giuliani Maurizio,
Cifone Maria Grazia
Publication year - 2015
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.24965
Subject(s) - liposuction , adipose tissue , in vitro , stem cell , microbiology and biotechnology , adipocyte , chemistry , biology , biochemistry , medicine , surgery
Nowadays, fat tissue transplantation is widely used in regenerative and reconstructive surgery. However, a shared method of lipoaspirate handling for ensuring a good quality fat transplant has not yet been established. The study was to identify a method to recover from the lipoaspirate samples the highest number of human viable adipose tissue‐derived stem cells (hADSCs) included in stromal vascular fraction (SVF) cells and of adipocytes suitable for transplantation, avoiding an extreme handling. We compared the lipoaspirate spontaneous stratification (10‐20‐30 min) with the centrifugation technique at different speeds (90‐400‐1500 × g). After each procedure, lipoaspirate was separated into top oily lipid layer, liquid fraction, “middle layer”, and bottom layer. We assessed the number of both adipocytes in the middle layer and SVF cells in all layers. The histology of middle layer and the surface phenotype of SVF cells by stemness markers (CD105+, CD90+, CD45−) was analyzed as well. The results showed a normal architecture in all conditions except for samples centrifuged at 1500 × g. In both methods, the flow cytometry analysis showed that greater number of ADSCs was in middle layer; in the fluid portion and in bottom layer was not revealed significant expression levels of stemness markers. Our findings indicate that spontaneous stratification at 20 min and centrifugation at 400 × g are efficient approaches to obtain highly viable ADSCs cells and adipocytes, ensuring a good thickness of lipoaspirate for autologous fat transfer. Since an important aspect of surgery practice consists of gain time, the 400 × g centrifugation could be the recommended method when the necessary instrumentation is available. J. Cell. Physiol. 230: 1974–1981, 2015. © 2015 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.