title: Ovine carotid artery-derived cells as an optimized supportive cell layer in 2-D capillary network assays.
abstract: BACKGROUND
  Endothelial cell co-culture assays are differentiation assays which simulate the formation of capillary-like tubules with the aid of a supportive cell layer. Different cell types have been employed as a supportive cell layer, including human pulmonary artery smooth muscle cells (PASMCs) and human mammary fibroblasts. However, these sources of human tissue-derived cells are limited, and more readily accessible human or animal tissue-derived cell sources would simplify the endothelial cell co-culture assay. In the present study, we investigated the potential use of alternative, accessible supportive cells for endothelial cell co-culture assay, including human umbilical cord and ovine carotid artery.
  
  
  RESULTS
  Human umbilical artery SMCs (HUASMCs) and ovine carotid artery-derived cells were seeded into 96-well plates, followed by addition of human umbilical vein endothelial cells (HUVECs). Nine days after co-culture, cells were fixed, immunostained and analysed using an in vitro angiogenesis quantification tool. Capillary-like structures were detected on ovine carotid artery-derived supportive cell layers. The initial cell number, as well as pro- and anti-angiogenic factors (VEGF, PDGF-BB and Bevacizumab), had a positive or negative influence on the number of capillary-like structures. Furthermore, HUVECs from different donors showed distinct levels of VEGF receptor-2, which correlated with the amount of capillary-like structures. In the case of HUASMC supportive cell layers, HUVECs detached almost completely from the surface.
  
  
  CONCLUSIONS
  Cells of different origin have a varying applicability regarding the endothelial cell co-culture assay: under the conditions described here, ovine carotid artery-derived cells seem to be more suitable than HUASMCs for an endothelial co-culture assay. Furthermore, the ovine carotid artery-derived cells are easier to obtain and are in more abundant supply than the currently used dermal or breast tissue cells. The use of ovine carotid artery-derived cells simplifies the endothelial co-culture assay with respect to testing large amounts of pro- and anti-angiogenic factors.
authors.affiliation: Department of Tissue Engineering & Textile Implants, AME - Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Weinandy
authors.affiliation: Department of Tissue Engineering & Textile Implants, AME - Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Babczyk
authors.affiliation: Department of Neurosurgery, Medical Faculty, RWTH Aachen University, Aachen, Germany.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Dreier
authors.affiliation: Institute of Pathology, University Medical Center, Johannes Gutenberg University of Mainz, Mainz, Germany.
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Unger
authors.affiliation: School of Medicine & Medical Science, Health Sciences Centre, University College Dublin, Dublin, Ireland.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Flanagan
authors.affiliation: Institute of Pathology, University Medical Center, Johannes Gutenberg University of Mainz, Mainz, Germany.
authors.forename: C James
authors.initials: CJ
authors.lastname: Kirkpatrick
authors.affiliation: Department of Cell Biology, Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Zenke
authors.affiliation: Institute of Technical and Macromolecular Chemistry (ITMC), RWTH Aachen University, Aachen, Germany.
authors.forename: Doris
authors.initials: D
authors.lastname: Klee
authors.affiliation: Department of Tissue Engineering & Textile Implants, AME - Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Jockenhoevel
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology; D000077214:Becaplermin; D000068258:Bevacizumab; D002196:Capillaries / Q000166:cytology*; D002339:Carotid Arteries / Q000166:cytology*; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D061252:Feeder Cells / Q000166:cytology*; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D012756:Sheep; D014470:Umbilical Cord / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D020574:Proto-Oncogene Proteins c-sis; D042461:Vascular Endothelial Growth Factor A; D000077214:Becaplermin; D000068258:Bevacizumab; D040301:Vascular Endothelial Growth Factor Receptor-2