Abstract
BackgroundDue to its low capacity for self-repair, articular cartilage is highly susceptible to damage and deterioration, which leads to the development of degenerative joint diseases such as osteoarthritis (OA). Menstrual blood-derived mesenchymal stem/stromal cells (MenSCs) are much less characterized, as compared to bone marrow mesenchymal stem/stromal cells (BMMSCs). However, MenSCs seem an attractive alternative to classical BMMSCs due to ease of access and broader differentiation capacity. The aim of this study was to evaluate chondrogenic differentiation potential of MenSCs and BMMSCs stimulated with transforming growth factor β (TGF-β3) and activin A.MethodsMenSCs (n = 6) and BMMSCs (n = 5) were isolated from different healthy donors. Expression of cell surface markers CD90, CD73, CD105, CD44, CD45, CD14, CD36, CD55, CD54, CD63, CD106, CD34, CD10, and Notch1 was analyzed by flow cytometry. Cell proliferation capacity was determined using CCK-8 proliferation kit and cell migration ability was evaluated by scratch assay. Adipogenic differentiation capacity was evaluated according to Oil-Red staining and osteogenic differentiation according to Alizarin Red staining. Chondrogenic differentiation (activin A and TGF-β3 stimulation) was investigated in vitro and in vivo (subcutaneous scaffolds in nude BALB/c mice) by expression of chondrogenic genes (collagen type II, aggrecan), GAG assay and histologically. Activin A protein production was evaluated by ELISA during chondrogenic differentiation in monolayer culture.ResultsMenSCs exhibited a higher proliferation rate, as compared to BMMSCs, and a different expression profile of several cell surface markers. Activin A stimulated collagen type II gene expression and glycosaminoglycan synthesis in TGF-β3 treated MenSCs but not in BMMSCs, both in vitro and in vivo, although the effects of TGF-β3 alone were more pronounced in BMMSCs in vitro.ConclusionThese data suggest that activin A exerts differential effects on the induction of chondrogenic differentiation in MenSCs vs. BMMSCs, which implies that different mechanisms of chondrogenic regulation are activated in these cells. Following further optimization of differentiation protocols and the choice of growth factors, potentially including activin A, MenSCs may turn out to be a promising population of stem cells for the development of cell-based therapies with the capacity to stimulate cartilage repair and regeneration in OA and related osteoarticular disorders.
Highlights
Human articular cartilage has a poor capacity for intrinsic repair and a weak ability to restore its lesions leading to the development of progressive and degenerative diseases such as osteoarthritis (OA), which is one of the most common forms of arthritis across the world [1]
Surface marker expression in mesenchymal stem/stromal cells (MenSCs) and bone marrow mesenchymal stem/stromal cells (BMMSCs) In order to determine and compare immunophenotypic profiles of MenSCs and BMMSCs, the cells were stained with antibodies against mesenchymal stem/ stromal cells (MSCs) surface markers (CD44, CD73, CD90, CD105), hematopoietic (CD14, CD34, CD36, CD45), and pluripotential stem cell markers (CD10, CD54, CD55, CD63, CD106, Notch1) and analyzed by flow cytometry (Fig. 1)
The results demonstrate that MenSCs and BMMSCs are able to differentiate into adipogenic and osteogenic lineage cells
Summary
Human articular cartilage has a poor capacity for intrinsic repair and a weak ability to restore its lesions leading to the development of progressive and degenerative diseases such as osteoarthritis (OA), which is one of the most common forms of arthritis across the world [1]. Menstrual blood is a unique body fluid, a renewable and sustainable source of multipotent stem cells (MenSCs) for regenerative medicine, which may originate from different subpopulations of endometrium, as reviewed [7, 8]. These cells are of great interest due to their ease of access, as their collection does not require any complicated procedures, permission of ethics authorities, or invasive surgical procedures [9, 10]. The aim of this study was to evaluate chondrogenic differentiation potential of MenSCs and BMMSCs stimulated with transforming growth factor β (TGFβ3) and activin A
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