title: EGF and hydrocortisone as critical factors for the co-culture of adipogenic differentiated ASCs and endothelial cells.
abstract: In vitro composed vascularized adipose tissue is and will continue to be in great demand e.g. for the treatment of extensive high-graded burns or the replacement of tissue after tumor removal. Up to date, the lack of adequate culture conditions, mainly a culture medium, decelerates further achievements. In our study, we evaluated the influence of epidermal growth factor (EGF) and hydrocortisone (HC), often supplemented in endothelial cell (EC) specific media, on the co-culture of adipogenic differentiated adipose-derived stem cells (ASCs) and microvascular endothelial cells (mvECs). In ASCs, EGF and HC are thought to inhibit adipogenic differentiation and have lipolytic activities. Our results showed that in indirect co-culture for 14 days, adipogenic differentiated ASCs further incorporated lipids and partly gained an univacuolar morphology when kept in media with low levels of EGF and HC. In media with high EGF and HC levels, cells did not incorporate further lipids, on the contrary, cells without lipid droplets appeared. Glycerol release, to measure lipolysis, also increased with elevated amounts of EGF and HC in the culture medium. Adipogenic differentiated ASCs were able to release leptin in all setups. MvECs were functional and expressed the cell specific markers, CD31 and von Willebrand factor (vWF), independent of the EGF and HC content as long as further EC specific factors were present. Taken together, our study demonstrates that adipogenic differentiated ASCs can be successfully co-cultured with mvECs in a culture medium containing low or no amounts of EGF and HC, as long as further endothelial cell and adipocyte specific factors are available.
authors.affiliation: Reutlingen University, Alteburgstr. 150, 72762 Reutlingen, Germany.
authors.forename: Ann-Cathrin
authors.initials: AC
authors.lastname: Volz
authors.affiliation: University of Stuttgart, Institute of Interfacial Process Engineering and Plasma Technology, Nobelstraße 12, 70569 Stuttgart, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Huber
authors.affiliation: Esslingen University of Applied Sciences, Kanalstraße 33, 73728 Esslingen, Germany.
authors.forename: Alina Maria
authors.initials: AM
authors.lastname: Schwandt
authors.affiliation: Reutlingen University, Alteburgstr. 150, 72762 Reutlingen, Germany; Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Nobelstraße 12, 70569 Stuttgart, Germany. Electronic address: petra.kluger@reutlingen-university.de.
authors.forename: Petra Juliane
authors.initials: PJ
authors.lastname: Kluger
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D050156:Adipogenesis; D000273:Adipose Tissue / Q000166:cytology*; D053687:Adult Stem Cells / Q000166:cytology* / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D017077:Culture Media, Conditioned / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D020738:Leptin / Q000378:metabolism; D066292:Lipid Droplets / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D020738:Leptin; D014841:von Willebrand Factor; D004815:Epidermal Growth Factor; D006854:Hydrocortisone
keywords: Adipocyte; Adipogenic differentiation; Adipose-derived stem cell (ASC); Co-culture; EGF; Endothelial cell; Hydrocortisone