Effect of reduced oxygen tension on chondrogenesis and osteogenesis in adipose-derived mesenchymal cells

Stanford University, Palo Alto, California, United States
AJP Cell Physiology (Impact Factor: 3.78). 05/2006; 290(4):C1139-46. DOI: 10.1152/ajpcell.00415.2005
Source: PubMed


Recent studies have demonstrated that adipose-derived mesenchymal cells (AMCs) offer great promise for cell-based therapies because of their ability to differentiate toward bone, cartilage, and fat. Given that cartilage is an avascular tissue and that mesenchymal cells experience hypoxia during prechondrogenic condensation in endochondral ossification, the goal of this study was to understand the influence of oxygen tension on AMC differentiation into bone and cartilage. In vitro chondrogenesis was induced using a three-dimensional micromass culture model supplemented with TGF-beta1. Collagen II production and extracellular matrix proteoglycans were assessed with immunohistochemistry and Alcian blue staining, respectively. Strikingly, micromasses differentiated in reduced oxygen tension (2% O(2)) showed markedly decreased chondrogenesis. Osteogenesis was induced using osteogenic medium supplemented with retinoic acid or vitamin D and was assessed with alkaline phosphatase activity and mineralization. AMCs differentiated in both 21 and 2% O(2) environments. However, osteogenesis was severely diminished in a low-oxygen environment. These data demonstrated that hypoxia strongly inhibits in vitro chondrogenesis and osteogenesis in AMCs.

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    • "The direct interaction between vascular endothelial cells and bone-forming osteoblasts leads to deposition of osteoid and begin of the mineralization process [19]. In cell culture experiments in vitro, a reduced oxygen tension has been accompanied by an increased proliferation rate and a decreased differentiation capacity of stem and osteoprogenitor cells compared to normoxic culture conditions [20] [21] [22] [23] [24] [25] [26]. Therefore, it is of great importance to monitor oxygen concentration inside bioreactors and even more inside scaffolds during the cultivation process. "
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    ABSTRACT: A three-dimensional computational fluid dynamics- (CFD-) model based on a differential pressure laminar flow bioreactor prototype was developed to further examine performance under changing culture conditions. Cell growth inside scaffolds was simulated by decreasing intrinsic permeability values and led to pressure build-up in the upper culture chamber. Pressure release by an integrated bypass system allowed continuation of culture. The specific shape of the bioreactor culture vessel supported a homogenous flow profile and mass flux at the scaffold level at various scaffold permeabilities. Experimental data showed an increase in oxygen concentration measured inside a collagen scaffold seeded with human mesenchymal stem cells when cultured in the perfusion bioreactor after 24 h compared to static culture in a Petri dish (dynamic: 11% O 2 versus static: 3% O 2 ). Computational fluid simulation can support design of bioreactor systems for tissue engineering application.
    08/2015; 2015(3):1-9. DOI:10.1155/2015/320280
    • " gamma and LPL is up - regulated ( Basciano et al . , 2011 ; Grayson et al . , 2007 ) . Equal MSC osteo - and adipo - potential has been demonstrated under both hypoxic and normoxic conditions ( Dos Santos et al . , 2010 ) . Incubation at low oxygen levels ( 2% ) has also been shown to inhibit the chondrogenic differentiation of murine AT - MSCs ( Malladi et al . , 2006 ) ."
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    ABSTRACT: Multipotent mesenchymal stromal cells (MSCs) are involved in the organization and maintenance of tissue integrity. MSCs have also attracted attention as a promising tool for cell therapy and regenerative medicine. However, their usage is limited due to cell impairment induced by an extremely harsh microenvironment during transplantation ex vivo. The microenvironment of MSCs in tissue depots is characterized by rather low oxygen consumption, demonstrating that MSCs might be quite resistant to oxygen limitation. However, accumulated data revealed that the response of MSCs to hypoxic conditions are rather controversial, demonstrating both damaging and ameliorating effects. Here, we make an attempt to summarize recent knowledge on the survival of MSCs under low oxygen conditions of various duration and severity, and to elucidate the mechanisms of MSC resistance/sensitivity to hypoxic impact.
    Mitochondrion 11/2014; 19. DOI:10.1016/j.mito.2014.07.005 · 3.25 Impact Factor
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    • "In addition, Lawyer, et al. [35] demonstrated that hypoxia stabilized the expression of HIF-1a in chondrocytes in comparison to normoxia . Given the fact that cartilage tissue is physiologically adapted to a lower oxygen tension compared to other tissues [23], we suggest that ASCs have a greater potential to undergo chondrogenic differentiation at oxygen levels as low as 2% O 2 than at higher oxygen tension. "
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    ABSTRACT: Adipose-derived stem cells (ASCs) have been found adapted to a specific niche with low oxygen tension (hypoxia) in the body. As an important component of this niche, oxygen tension has been known to play a critical role in the maintenance of stem cell characteristics. However, the effect of O2 tension on their functional properties has not been well determined. In this study, we investigated the effects of O2 tension on ASCs stemness, differentiation and proliferation ability. Human ASCs were cultured under normoxia (21% O2) and hypoxia (2% O2). We found that hypoxia increased ASC stemness marker expression and proliferation rate without altering their morphology and surface markers. Low oxygen tension further enhances the chondrogenic differentiation ability, but reduces both adipogenic and osteogenic differentiation potential. These results might be correlated with the increased expression of HIF-1α under hypoxia. Taken together, we suggest that growing ASCs under 2% O2 tension may be important in expanding ASCs effectively while maintaining their functional properties for clinical therapy, particularly for the treatment of cartilage defects.
    Biochemical and Biophysical Research Communications 05/2014; 448(2). DOI:10.1016/j.bbrc.2014.04.096 · 2.30 Impact Factor
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