Allogeneic Serum and Macromolecular Crowding Maintain Native Equine Tenocyte Function in Culture

Abstract

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.

Macromolecular crowding in human tenocyte and skin fibroblast cultures: A comparative analysis

“…Cryopreserved hDFs (PCS-201-012) were obtained from ATCC (UK). hTCs were isolated from surgical remains of healthy tendons donated by patients (appropriate ethical approvals and informed consents were in place) undergoing tendon-related procedures at Merlin Park University Hospital (Ireland) and Galway University Hospital (Ireland), by means of an explant tissue culture approach [ 45 ]. Three different donors were used for this study.…”