title: Adipose tissue derived stem cells: in vitro and in vivo analysis of a standard and three commercially available cell-assisted lipotransfer techniques.
abstract: BACKGROUND
  Autologous fat grafting is commonly used to correct soft-tissue contour deformities. However, results are impaired by a variable and unpredictable resorption rate. Autologous adipose-derived stromal cells in combination with lipoinjection (cell-assisted lipotransfer) seem to favor a long-term persistence of fat grafts, thus fostering the development of devices to be used in the operating room at the point of care, to isolate the stromal vascular fraction (SVF) and produce SVF-enhanced fat grafts with safe and standardized protocols. Focusing on patients undergoing breast reconstruction by lipostructure, we analyzed a standard technique, a modification of the Coleman's procedure, and three different commercially available devices (Lipokit, Cytori, Fastem), in terms of 1) ability to enrich fat grafts in stem cells and 2) clinical outcome at 6 and 12 months.
  
  
  METHODS
  To evaluate the ability to enrich stem cells, we compared, for each patient (n=20), the standard lipoaspirate with the respective stem cell-enriched one, analyzing yield, immunophenotype and colony-forming capacity of the SVF cells as well as immunophenotype, clonogenicity and multipotency of the obtained adipose stem cells (ASCs). Regarding the clinical outcome, we compared, by ultrasonography imaging, changes at 6 and 12 months in the subcutaneous thickness of patients treated with stem-cell enriched (n=14) and standard lipoaspirates (n=16).
  
  
  RESULTS
  Both methods relying on the enzymatic isolation of primitive cells led to significant increase in the frequency, in the fat grafts, of SVF cells as well as of clonogenic and multipotent ASCs, while the enrichment was less prominent for the device based on the mechanical isolation of the SVF. From a clinical point of view, patients treated with SVF-enhanced fat grafts demonstrated, at six months, a significant superior gain of thickness of both the central and superior-medial quadrants with respect to patients treated with standard lipotransfer. In the median-median quadrant the effect was still persistent at 12 months, confirming an advantage of lipotransfer technique in enriching improving long-term fat grafts.
  
  
  CONCLUSIONS
  This comparative study, based on reproducible biological and clinical parameters and endpoints, showed an advantage of lipotransfer technique in enriching fat grafts in stem cells and in favoring, clinically, long-term fat grafts.
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. rossana.domenis@uniud.it.
authors.forename: Rossana
authors.initials: R
authors.lastname: Domenis
authors.affiliation: Clinic of Plastic and Reconstructive Surgery of Udine, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. laralazzaro@hotmail.com.
authors.forename: Lara
authors.initials: L
authors.lastname: Lazzaro
authors.affiliation: Clinic of Plastic and Reconstructive Surgery of Udine, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. sarah2012@hotmail.com.
authors.forename: Sarah
authors.initials: S
authors.lastname: Calabrese
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. damianomng@gmail.com.
authors.forename: Damiano
authors.initials: D
authors.lastname: Mangoni
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. byoteck@libero.it.
authors.forename: Annarita
authors.initials: A
authors.lastname: Gallelli
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. jenny_bourkoula@hotmail.com.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Bourkoula
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. ivana.manini@tiscali.it.
authors.forename: Ivana
authors.initials: I
authors.lastname: Manini
authors.affiliation: Azienda Ospedaliero-Universitaria of Udine, P.le S. Maria della Misericordia 15, 33100, Udine, Italy. nataschab@libero.it.
authors.forename: Natascha
authors.initials: N
authors.lastname: Bergamin
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. t_barbara2001@yahoo.it.
authors.forename: Barbara
authors.initials: B
authors.lastname: Toffoletto
authors.affiliation: Azienda Ospedaliero-Universitaria of Udine, P.le S. Maria della Misericordia 15, 33100, Udine, Italy. carloalberto.beltrami@uniud.it.
authors.forename: Carlo A
authors.initials: CA
authors.lastname: Beltrami
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. antonio.beltrami@uniud.it.
authors.forename: Antonio P
authors.initials: AP
authors.lastname: Beltrami
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. daniela.cesselli@uniud.it.
authors.forename: Daniela
authors.initials: D
authors.lastname: Cesselli
authors.affiliation: Clinic of Plastic and Reconstructive Surgery of Udine, University of Udine, P.le Kolbe 4, 33100, Udine, Italy. piercamillo.parodi@uniud.it.|Azienda Ospedaliero-Universitaria of Udine, P.le S. Maria della Misericordia 15, 33100, Udine, Italy. piercamillo.parodi@uniud.it.
authors.forename: Pier Camillo
authors.initials: PC
authors.lastname: Parodi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D000368:Aged; D000954:Antigens, Surface / Q000378:metabolism; D001940:Breast / Q000601:surgery; D002454:Cell Differentiation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D015187:Lipectomy; D008408:Mastectomy; D008875:Middle Aged; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D016217:Ultrasonography, Mammary; D055815:Young Adult
chemical_list: D000954:Antigens, Surface