title: Intraoperative use of enriched collagen and elastin matrices with freshly isolated adipose-derived stem/stromal cells: a potential clinical approach for soft tissue reconstruction.
abstract: BACKGROUND
  Adipose tissue contains a large number of multipotent cells, which are essential for stem cell-based therapies. The combination of this therapy with suitable commercial clinically used matrices, such as collagen and elastin matrices (i.e. dermal matrices), is a promising approach for soft tissue reconstruction. We previously demonstrated that the liposuction method affects the adherence behaviour of freshly isolated adipose-derived stem/stromal cells (ASCs) on collagen and elastin matrices. However, it remains unclear whether freshly isolated and uncultured ASCs could be directly transferred to matrices during a single transplantation operation without additional cell culture steps.
  
  
  METHODS
  After each fat harvesting procedure, ASCs were isolated and directly seeded onto collagen and elastin matrices. Different time intervals (i.e. 1, 3 and 24 h) were investigated to determine the time interval needed for cellular attachment to the collagen and elastin matrices. Resazurin-based vitality assays were performed after seeding the cells onto the collagen and elastin matrices. In addition, the adhesion and migration of ASCs on the collagen and elastin matrices were visualised using histology and two-photon microscopy.
  
  
  RESULTS
  A time-dependent increase in the number of viable ASCs attached to the collagen and elastin matrices was observed. This finding was supported by mitochondrial activity and histology results. Importantly, the ASCs attached and adhered to the collagen and elastin matrices after only 1 h of ex vivo enrichment. This finding was also supported by two-photon microscopy, which revealed the presence and attachment of viable cells on the upper layer of the construct.
  
  
  CONCLUSIONS
  Freshly isolated uncultured ASCs can be safely seeded onto collagen and elastin matrices for ex vivo cellular enrichment of these constructs after liposuction. Although we observed a significant number of seeded cells on the matrices after a 3-h enrichment time, we also observed an adequate number of isolated cells after a 1-h enrichment time. However, this approach must be optimised for clinical use. Thus, in vivo studies and clinical trials are needed to investigate the feasibility of this approach.
authors.affiliation: Department of Plastic, Reconstructive and Hand Surgery - Burn Center, Medical Faculty, RWTH Aachen University, Pauwelsstr, 30, Aachen D-52074, Germany. zalharbi@ukaachen.de.
authors.forename: Ziyad
authors.initials: Z
authors.lastname: Alharbi
authors.forename: Sultan
authors.initials: S
authors.lastname: Almakadi
authors.forename: Christian
authors.initials: C
authors.lastname: Opländer
authors.forename: Michael
authors.initials: M
authors.lastname: Vogt
authors.forename: Hans-Oliver
authors.initials: HO
authors.lastname: Rennekampff
authors.forename: Norbert
authors.initials: N
authors.lastname: Pallua
mesh_terms: D062088:Acellular Dermis; D000328:Adult; D002448:Cell Adhesion; D002465:Cell Movement; D002470:Cell Survival; D003094:Collagen; D004549:Elastin; D005260:Female; D006801:Humans; D007432:Intraoperative Period; D015187:Lipectomy; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008875:Middle Aged; D019651:Plastic Surgery Procedures / Q000379:methods; D017695:Soft Tissue Injuries / Q000601:surgery; D050151:Subcutaneous Fat / Q000166:cytology*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D003094:Collagen; D004549:Elastin