title: Effect of Cell Seeding Density and Inflammatory Cytokines on Adipose Tissue-Derived Stem Cells: an in Vitro Study.
abstract: Adipose tissue-derived stem cells (ASCs) are known to be able to promote repair of injured tissue via paracrine factors. However, the effect of cell density and inflammatory cytokines on the paracrine ability of ASCs remains largely unknown. To investigate these effects, ASCs were cultured in 8000 cells/cm2, 20,000 cells/cm2, 50,000 cells/cm2, and 400,000 cells/cm2with and without 10 or 20 ng/ml tumor necrosis factor alpha (TNFα) and 25 or 50 ng/ml interferon gamma (IFNγ). ASC-sheets formed at 400,000 cells/cm2after 48 h of culture. With increasing concentrations of TNFα and IFNγ, ASC-sheets with 400,000 cells/cm2had increased production of angiogenic factors Vascular Endothelial Growth Factor and Fibroblast Growth Factor and decreased expression of pro-inflammatory genes TNFA and Prostaglandin Synthase 2 (PTGS2) compared to lower density ASCs. Moreover, the conditioned medium of ASC-sheets with 400,000 cells/cm2stimulated with the low concentration of TNFα and IFNγ enhanced endothelial cell proliferation and fibroblast migration. These results suggest that a high cell density enhances ASC paracrine function might beneficial for wound repair, especially in pro-inflammatory conditions.
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.|Department of Otorhinolaryngology, Erasmus MC University Medical Center, Rotterdam, The Netherlands.|Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom, Thailand.
authors.forename: Panithi
authors.identifier: 0000-0002-6524-4765
authors.initials: P
authors.lastname: Sukho
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.|Hills Pet Nutrition Inc, Topeka, Kansas, USA.
authors.forename: Jolle
authors.initials: J
authors.lastname: Kirpensteijn
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Jan Willem
authors.initials: JW
authors.lastname: Hesselink
authors.affiliation: Department of Otorhinolaryngology, Erasmus MC University Medical Center, Rotterdam, The Netherlands.|Department of Orthopaedics, Erasmus MC University Medical Center, Wytemaweg 80, 3015, CN, Rotterdam, The Netherlands.
authors.forename: Gerjo J V M
authors.initials: GJ
authors.lastname: van Osch
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.|Department of Orthopaedics, Erasmus MC University Medical Center, Wytemaweg 80, 3015, CN, Rotterdam, The Netherlands.
authors.forename: Femke
authors.initials: F
authors.lastname: Verseijden
authors.affiliation: Department of Orthopaedics, Erasmus MC University Medical Center, Wytemaweg 80, 3015, CN, Rotterdam, The Netherlands. y.bastiaansen@erasmusmc.nl.
authors.forename: Yvonne M
authors.initials: YM
authors.lastname: Bastiaansen-Jenniskens
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D008875:Middle Aged; D019899:Paracrine Communication / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
chemical_list: D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D007371:Interferon-gamma; D051546:Cyclooxygenase 2
keywords: Adipose tissue-derived stem cell; Cell sheet; Inflammatory cytokine and paracrine ability; Seeding density