Allogeneic Serum and Macromolecular Crowding Maintain Native Equine Tenocyte Function in Culture
Andrea Rampin,
Ioannis Skoufos,
Michael Raghunath,
Athina Tzora,
Nikolaos Diakakis,
Nikitas Prassinos,
Dimitrios I. Zeugolis
Affiliations
Andrea Rampin
Laboratory of Animal Science, Nutrition and Biotechnology, School of Agriculture, University of Ioannina, 47100 Arta, Greece
Ioannis Skoufos
Laboratory of Animal Science, Nutrition and Biotechnology, School of Agriculture, University of Ioannina, 47100 Arta, Greece
Michael Raghunath
Center for Cell Biology and Tissue Engineering, Institute for Chemistry and Biotechnology, Zurich University of Applied Sciences, 8820 Wädenswil, Switzerland
Athina Tzora
Laboratory of Animal Science, Nutrition and Biotechnology, School of Agriculture, University of Ioannina, 47100 Arta, Greece
Nikolaos Diakakis
School of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
Nikitas Prassinos
School of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
Dimitrios I. Zeugolis
Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Charles Institute of Dermatology, Conway Institute of Biomolecular & Biomedical Research, School of Mechanical & Materials Engineering, University College Dublin (UCD), D04 V1W8 Dublin, Ireland
The absence of a native extracellular matrix and the use of xenogeneic sera are often associated with rapid tenocyte function losses during in vitro culture. Herein, we assessed the influence of different sera (equine serum and foetal bovine serum) on equine tenocyte morphology, viability, metabolic activity, proliferation and protein synthesis as a function of tissue-specific extracellular matrix deposition (induced via macromolecular crowding), aging (passages 3, 6, 9) and time in culture (days 3, 5, 7). In comparison to cells at passage 3, at day 3, in foetal bovine serum and without macromolecular crowding (traditional equine tenocyte culture), the highest number of significantly decreased readouts were observed for cells in foetal bovine serum, at passage 3, at day 5 and day 7 and without macromolecular crowding. Again, in comparison to traditional equine tenocyte culture, the highest number of significantly increased readouts were observed for cells in equine serum, at passage 3 and passage 6, at day 7 and with macromolecular crowding. Our data advocate the use of an allogeneic serum and tissue-specific extracellular matrix for effective expansion of equine tenocytes.
