Abstract
While both induction culture media and matrix have been reported to regulate the stem cell fate, little is known about which factor plays a more decisive role in directing the MSC differentiation lineage as well as the underlying mechanisms. To this aim, we seeded MSCs on HA-collagen and HA-synthetic hydrogel matrixes, which had demonstrated highly different potentials toward osteoblastic and chondrocytic differentiation lineages, respectively, and cultured them with osteogenic, chondrogenic and normal culture media, respectively. A systematic comparison has been carried out on the effects of induction media and matrix on MSC adhesion, cytoskeleton organization, proliferation, and in particular differentiation into the osteoblastic and chondrocytic lineages. The results demonstrated that the matrix selection had a much more profound effect on directing the differentiation lineage than the induction media did. The strong modulation effect on the transcription activities might be the critical factor contributing to the above observations in our study, where canonical Wnt-β-Catenin signal pathway was directly involved in the matrix-driven osteoblastic differentiation. Such findings not only provide a critical insight on natural cellular events leading to the osteoblastic and chondrocytic differentiations, but also have important implications in biomaterial design for tissue engineering applications.
Highlights
Mesenchymal stem cells (MSCs) are multipotent cells, present in bone marrow, that are able to differentiate into various anchoragedependent cell types, including neurons, myoblasts, osteoblasts, chondrocytes and so on [1, 2]
The alkaline phosphates (ALP) staining confirmed the striking modulating effect of matrix on osteoblastic differentiation, showing significant higher staining for HA-collagen compared with the HA-synthetic hydrogel, especially under the normal and osteogenic culture conditions (Fig. 1c)
The ALP activities were significantly suppressed under chondrogenic induction media for both matrices (Fig. 1a3)
Summary
Mesenchymal stem cells (MSCs) are multipotent cells, present in bone marrow, that are able to differentiate into various anchoragedependent cell types, including neurons, myoblasts, osteoblasts, chondrocytes and so on [1, 2]. Due to their capacity to differentiate into cells of connective tissue lineages, MSCs are promising candidates for use in cell-based therapies and tissue engineering applications. Directing the MSC differentiation into its desired lineage is critical for tissue regeneration and has drawn continued interests from the researchers. The usage of induction culture media is a common and effective method to direct the MSC differentiation toward the osteogenic, chondrogenic, adipogenic lineages and so forth [11–15]. The osteogenic induction medium, containing dexamethasone (Dex), b-glycerophosphate and ascorbic acid, has been generally used to induce the MSC differentiation into osteoblast, by increasing the alkaline phosphatase activity and accelerating bone mineralization [16–18]
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