Abstract
BackgroundHuman multipotent mesenchymal stromal cells (MSC) can be isolated from various tissues including bone marrow. Here, MSC participate as bone lining cells in the formation of the hematopoietic stem cell niche. In this compartment, the oxygen tension is low and oxygen partial pressure is estimated to range from 1% to 7%. We analyzed the effect of low oxygen tensions on human MSC cultured with platelet-lysate supplemented media and assessed proliferation, morphology, chromosomal stability, immunophenotype and plasticity.ResultsAfter transferring MSC from atmospheric oxygen levels of 21% to 1%, HIF-1α expression was induced, indicating efficient oxygen reduction. Simultaneously, MSC exhibited a significantly different morphology with shorter extensions and broader cell bodies. MSC did not proliferate as rapidly as under 21% oxygen and accumulated in G1 phase. The immunophenotype, however, was unaffected. Hypoxic stress as well as free oxygen radicals may affect chromosomal stability. However, no chromosomal abnormalities in human MSC under either culture condition were detected using high-resolution matrix-based comparative genomic hybridization. Reduced oxygen tension severely impaired adipogenic and osteogenic differentiation of human MSC. Elevation of oxygen from 1% to 3% restored osteogenic differentiation.ConclusionPhysiologic oxygen tension during in vitro culture of human MSC slows down cell cycle progression and differentiation. Under physiological conditions this may keep a proportion of MSC in a resting state. Further studies are needed to analyze these aspects of MSC in tissue regeneration.
Highlights
Human multipotent mesenchymal stromal cells (MSC) can be isolated from various tissues including bone marrow
As the cells adhered to plastic, the nonadherent hematopoietic cells in the culture were removed during changes of medium and virtually homogenous cultures with the characteristic confluent, spindle-shaped MSC were obtained after three passages
Effect of reduced oxygen concentrations on MSC immunophenotype The immunophenotype of human MSC is characterized by the cell surface expression of CD73, CD90, CD105, CD106, CD146 and MHC class I
Summary
Human multipotent mesenchymal stromal cells (MSC) can be isolated from various tissues including bone marrow. MSC participate as bone lining cells in the formation of the hematopoietic stem cell niche. In this compartment, the oxygen tension is low and oxygen partial pressure is estimated to range from 1% to 7%. Human multipotent mesenchymal stromal cells (MSC) obtained from bone marrow are characterized by a multilineage differentiation potential and a high proliferative capacity without losing their genetic stability [1]. The treatment of pediatric patients suffering from osteogenesis imperfecta was the first successful clinical application of MSC in regenerative medicine [3,7]. Regenerative properties of MSC could be demonstrated in the treatment of ischemic cardiovascular diseases [9]
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