title: Human infrapatellar fat pad-derived stem cells express the pericyte marker 3G5 and show enhanced chondrogenesis after expansion in fibroblast growth factor-2.
abstract: BACKGROUND
  Infrapatellar fat pad (IPFP) is a possible source of stem cells for the repair of articular cartilage defects. In this study, adherent proliferative cells were isolated from digests of IPFP tissue. The effects of the expansion of these cells in fibroblast growth factor-2 (FGF-2) were tested on their proliferation, characterisation, and chondrogenic potential.
  
  
  METHODS
  IPFP tissue was obtained from six patients undergoing total knee replacement, and sections were stained with 3G5, alpha smooth muscle actin, and von Willebrand factor to identify different cell types in the vasculature. Cells were isolated from IPFP, and both mixed populations and clonal lines derived from them were characterised for cell surface epitopes, including 3G5. Cells were expanded with and without FGF-2 and were tested for chondrogenic differentiation in cell aggregate cultures.
  
  
  RESULTS
  3G5-positive cells were present in perivascular regions in tissue sections of the IPFP, and proliferative adherent cells isolated from the IPFP were also 3G5-positive. However, 3G5 expression was on only a small proportion of cells in all populations and at all passages, including the clonally expanded cells. The cells showed cell surface epitope expression similar to adult stem cells. They stained strongly for CD13, CD29, CD44, CD90, and CD105 and were negative for CD34 and CD56 but were also negative for LNGFR (low-affinity nerve growth factor receptor) and STRO1. The IPFP-derived cells showed chondrogenic differentiation in cell aggregate cultures, and prior expansion with FGF-2 enhanced chondrogenesis. Expansion in FGF-2 resulted in greater downregulation of many cartilage-associated genes, but on subsequent chondrogenic differentiation, they showed stronger upregulation of these genes and this resulted in greater matrix production per cell.
  
  
  CONCLUSIONS
  These results show that these cells express mesenchymal stem cell markers, but further work is needed to determine the true origin of these cells. These results suggest that the expansion of these cells with FGF-2 has important consequences for facilitating their chondrogenic differentiation.
authors.affiliation: Faculty of Life Sciences, United Kingdom Centre for Tissue Engineering at the Wellcome Trust Centre for Cell Matrix Research, Michael Smith Building, University of Manchester, Oxford Road, Manchester, UK. wasimkhan@doctors.org.uk
authors.forename: Wasim S
authors.initials: WS
authors.lastname: Khan
authors.forename: Simon R
authors.initials: SR
authors.lastname: Tew
authors.forename: Adetola B
authors.initials: AB
authors.lastname: Adesida
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Hardingham
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010329:Patella / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020286:Pericytes / Q000166:cytology; D012216:Rheumatic Diseases / Q000628:therapy
chemical_list: D000911:Antibodies, Monoclonal; D006133:Growth Substances; D016222:Fibroblast Growth Factor 2