Human vascular model with defined stimulation medium - a characterization study.
The formation of blood vessels is a vital process in embryonic development and in normal physiology. Current vascular modelling is mainly based on animal biology leading to species-to-species variation when extrapolating the results to humans. Although there are a few human cell based vascular models available these assays are insufficiently characterized in terms of culture conditions and developmental stage of vascular structures. Therefore, well characterized vascular models with human relevance are needed for basic research, embryotoxicity testing, development of therapeutic strategies and for tissue engineering. We have previously shown that the in vitro vascular model based on co-culture of human adipose stromal cells (hASC) and human umbilical vein endothelial cells (HUVEC) is able to induce an extensive vascular-like network with high reproducibility. In this work we developed a defined serum-free vascular stimulation medium (VSM) and performed further characterization in terms of cell identity, maturation and structure to obtain a thoroughly characterized in vitro vascular model to replace or reduce corresponding animal experiments. The results showed that the novel vascular stimulation medium induced intact and evenly distributed vascular-like network with morphology of mature vessels. Electron microscopic analysis assured the three-dimensional microstructure of the network containing lumen. Additionally, elevated expressions of the main human angiogenesis-related genes were detected. In conclusion, with the new defined medium the vascular model can be utilized as a characterized test system for chemical testing as well as in creating vascularized tissue models.
FICAM, Finnish Centre for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.
Outi
O
Huttala
FICAM, Finnish Centre for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.
Hanna
H
Vuorenpää
FICAM, Finnish Centre for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.
Tarja
T
Toimela
Department of Obstetrics and Gynecology, Tampere University Hospital, Tampere, Finland.
Jukka
J
Uotila
Department of Plastic Surgery, Tampere University Hospital, Tampere, Finland.
Hannu
H
Kuokkanen
FICAM, Finnish Centre for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.|Department of Cell Biology, School of Medicine, University of Tampere, Tampere, Finland.
Timo
T
Ylikomi
FICAM, Finnish Centre for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.|Department of Cell Biology, School of Medicine, University of Tampere, Tampere, Finland.
Jertta-Riina
JR
Sarkanen
FICAM, Finnish Centre for Alternative Methods, School of Medicine, University of Tampere, Tampere, Finland.
Tuula
T
Heinonen
D000273:Adipose Tissue / Q000098:blood supply / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D003470:Culture Media / Q000737:chemistry*; D005434:Flow Cytometry; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology; D008854:Microscopy, Electron; D018919:Neovascularization, Physiologic / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D023822:Tissue Engineering; D018675:Toxicity Tests
D003470:Culture Media
angiogenesis; coculture techniques; mesenchymal stromal cells; serum-free media