A. Ritthaphai (Bristol, GB)
University of Bristol School of Cellular and Molecular MedicinePresenter Of 1 Presentation
P224 - The Impact of Hypoxia on Mesenchymal Stem Cells Isolated from Osteoarthritis Patients
Abstract
Purpose
Improving the outcomes for cartilage tissue engineering using mesenchymal stem cells (MSCs) requires in-depth understanding of their physiological environment in health and disease. Local oxygen tension has been implicated in regulating the physiology of MSCs with debatable results in the literature. In this study we focused on investigating the impact of hypoxia on the expansion and chondrogenic differentiation of MSCs isolated from osteoarthritis (OA) patients.
Methods and Materials
Adult MSCs were isolated from the bone marrow of patients with advanced OA (n=6) and incubated under normoxia (20% O2) and hypoxia (5% and 2% O2) for up to 18 days. Expanded cells were assessed for cell number and population doubling. 3D pellets were created from all expanded groups and cultured in chondrogenic medium under the same conditions for 7, 14, and 21 days. Harvested pellets were anlaysed by histology and qPCR for chondrogenic markers and hypoxia-induced factor (HIF)-1 and -2.
Results
MSCs demonstrated exponential growth for all groups until day 12 and plateaued afterwards. Cells grown in 5% O2 had the fastest growth rate followed by normoxia and 2% O2. There was gradual increase in doubling time for all groups. HIF-1 and -2 gene expression showed cyclical expression over time in hypoxic conditions compared to a stable expression in normoxia. 3D pellets exhibited heterogeneity among patients in pellet size and chondrogenic markers expression without measured difference among oxygenation states. HIF-1 and -2 expression however, correlated with the level of hypoxia.
Conclusion
MSCs isolated from OA patients are less sensitive to changes in oxygen tension during expansion and chondrogenic differentiation. The inherent heterogeneity associated with MSCs from this patient group could be a confounding factor. A study comparing MSCs from healthy and OA patients is warranted.
Presenter Of 1 Presentation
P224 - The Impact of Hypoxia on Mesenchymal Stem Cells Isolated from Osteoarthritis Patients
Abstract
Purpose
Improving the outcomes for cartilage tissue engineering using mesenchymal stem cells (MSCs) requires in-depth understanding of their physiological environment in health and disease. Local oxygen tension has been implicated in regulating the physiology of MSCs with debatable results in the literature. In this study we focused on investigating the impact of hypoxia on the expansion and chondrogenic differentiation of MSCs isolated from osteoarthritis (OA) patients.
Methods and Materials
Adult MSCs were isolated from the bone marrow of patients with advanced OA (n=6) and incubated under normoxia (20% O2) and hypoxia (5% and 2% O2) for up to 18 days. Expanded cells were assessed for cell number and population doubling. 3D pellets were created from all expanded groups and cultured in chondrogenic medium under the same conditions for 7, 14, and 21 days. Harvested pellets were anlaysed by histology and qPCR for chondrogenic markers and hypoxia-induced factor (HIF)-1 and -2.
Results
MSCs demonstrated exponential growth for all groups until day 12 and plateaued afterwards. Cells grown in 5% O2 had the fastest growth rate followed by normoxia and 2% O2. There was gradual increase in doubling time for all groups. HIF-1 and -2 gene expression showed cyclical expression over time in hypoxic conditions compared to a stable expression in normoxia. 3D pellets exhibited heterogeneity among patients in pellet size and chondrogenic markers expression without measured difference among oxygenation states. HIF-1 and -2 expression however, correlated with the level of hypoxia.
Conclusion
MSCs isolated from OA patients are less sensitive to changes in oxygen tension during expansion and chondrogenic differentiation. The inherent heterogeneity associated with MSCs from this patient group could be a confounding factor. A study comparing MSCs from healthy and OA patients is warranted.