title: A non-enzymatic method for isolating human adipose tissue-derived stromal stem cells.
abstract: OBJECTIVE
  The isolation of human adipose stromal/stem cells (ASCs) currently relies on the use of the enzyme collagenase, which digests the triple helix region of peptide bonds in the collagen of adipose tissue. Collagenase is an expensive reagent derived from a bacterial source, and its use in isolating ASCs is a time-consuming procedure. This experiment evaluated the extraction of ASCs without an enzymatic digest.
  
  
  METHODS
  We used a simple method of washing adipose tissue to isolate and characterize the cells and compared this method with the enzymatic procedure in terms of processing time, stem cell yield, differentiation potential and immunophenotype.
  
  
  RESULTS
  Based on fluorescence activated cell sorting analysis, the stromal vascular fractions isolated with the washing method displayed a distinct and potentially favorable immunophenotype relative to the collagenase digestion. This difference may reflect the absence of chemical alteration of the cells by collagenase digestion. Independent of the isolation procedure, the resulting passaged ASCs were comparable based on immunophenotype and adipogenic and osteogenic differentiation potential.
  
  
  CONCLUSIONS
  Although using collagenase substantially increases cell yield, the two methods yield a similar cell product.
authors.affiliation: Stem Cell Biology Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA.
authors.forename: Forum S
authors.initials: FS
authors.lastname: Shah
authors.forename: Xiying
authors.initials: X
authors.lastname: Wu
authors.forename: Marilyn
authors.initials: M
authors.lastname: Dietrich
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rood
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Gimble
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases / Q000737:chemistry; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis; D023822:Tissue Engineering
chemical_list: D017364:Collagenases
keywords: adipose stromal/stem cells; collagenase; lipoaspirate; non-enzymatic; tissue engineering