Human mesenchymal stem cells: Influence of oxygen pressure on proliferation and chondrogenic differentiation in fibrin glue in vitro

Abstract

Tissue engineering using biomaterials is a promising solution for cartilage replacement. The purpose of this study was to investigate whether the fibrin sealant Tissucol(R) provides a suitable scaffold for re-implanting stem cells during chondrogenic replacement therapy. Pluripotent stem cells were isolated from adult human bone marrow (hMSCs), cultured and characterized by FACS (CD105+/CD106+, CD45-/CD14-/CD34-). A large-holed porous hMSC-containing fibrin matrix was built that allowed hMSCs to survive throughout the period of culture (42 days) in either proliferation or chondrogenic differentiation medium under normoxic (21% O2) or hypoxic (3% O2) conditions. Morphology (as determined by electron microscopy) and proliferation (Ki67 staining) of the embedded hMSCs did not markedly vary under normoxic and hypoxic culture even after 42 days in culture. The stem cell marker Oct-4 was expressed during the whole culture period. Under chondrogenic differentiation conditions, especially under hypoxic conditions, we observed rounded chondrocyte-like cell types and a chondral phenotype assessed by mRNA expression of collagen II and Alcian blue staining. hMSCs seeded into large-holed porous preparations of Tissucol survive, proliferate and keep their stem cell character. Furthermore, culturing the cells in a corresponding medium induces chondrogenic differentiation, which could be remarkably and significantly enhanced under hypoxic conditions.

Role of hypoxia and growth and differentiation factor-5 on differentiation of human mesenchymal stem cells towards intervertebral nucleus pulposus-like cells

“…Treatment with GDF5/Dex showed similar effects, although with different collagen to aggrecan ratios, while hypoxic conditions generally enhanced the chondrogenic response. These benefi cial effects of low oxygen tension in terms of chondrogenic or NP-like differentiation of MSCs are in agreement with previously described data and are likely to refl ect the in vivo situation that is characterised by physiologically low oxygen concentration in NP and cartilaginous tissues (Risbud et al, 2004;Baumgartner et al, 2010;Markway et al, 2010).…”

“…In the present study we measured the influence of hypoxia on human bone marrow derived MSCs in three dimensional alginate cultures as a modulator of the effects of transforming growth factor beta 1 (TGFβ1), GDF5 and coculture with bovine nucleus pulposus cells (bNPC), for the purpose of inducing an IVD-like phenotype. Hypoxic conditions applying 2 % oxygen tension were chosen to mimic the avascular hypoxic environment of the NP tissue; previous studies had shown enhanced chondrogenic and NP-like differentiation in MSCs cultured under 2-3 % oxygen (Risbud et al, 2004;Felka et al, 2009;Baumgartner et al, 2010;Markway et al, 2010). We hypothesised that different combinations of hypoxia with growth factors or coculture may have synergistic or modulatory effects and some of these combinations may result in a differentiation pattern approaching that of IVD cells.…”

The effects of dynamic loading on the intervertebral disc

“…Furthermore, the high volumetric stress environment of the IVD could direct MSC differentiation towards a chondrogenic phenotype [16,94]. Hypoxia has been shown to impair adipogenesis and osteogenesis of human MSCs [38], and as a potent chondrogenesis promoter than dynamic compression or hydrostatic pressure in the presence of transforming growth factor b (TGF-b) [7,74,77,105]. Nevertheless, low oxygen tension and hydrostatic pressure alone without TGF-b could not initiate chondrogenesis of MSC [105].…”

Human Wharton's Jelly Mesenchymal Stem Cells Maintain the Expression of Key Immunomodulatory Molecules When Subjected to Osteogenic, Adipogenic and Chondrogenic Differentiation In Vitro: New Perspectives for Cellular Therapy

“…Particular interest was demonstrated in the use of specific media to support proliferation by MSCs and their transdifferentiation process. Some studies showed that hypoxia can induce chondrogenesis and chondrogenic differentiation of MSCs [62][63][64]. Further reports highlighted the use of a bioreactor with co-culture of MSCs and other stem cells to study the culture and mechanical stimulation [65].…”