Screening for Growth-Factor Combinations Enabling Synergistic Differentiation of Human MSC to Nucleus Pulposus Cell-Like Cells

Background: Multiple studies have examined the potential of growth factors (GF) to enable mesenchymal stromal cells (MSC) to nucleus pulposus (NP) cell-like cell differentiation. Here we screened a wide range of GF and GF combinations for supporting NP cell-like cell differentiation. Methods: Human...

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Bibliographic Details
Published in:Applied sciences 2021-04, Vol.11 (8), p.3673
Main Authors: Morita, Kosuke, Schol, Jordy, Volleman, Tibo N. E., Sakai, Daisuke, Sato, Masato, Watanabe, Masahiko
Format: Article
Language:English
Subjects:
Cell differentiation
Collagen
Degenerative disc disease
differentiation
Differentiation (biology)
Glycosaminoglycans
Growth differentiation factor 5
growth factor
Growth factors
Intervertebral discs
Markers
mesenchymal stromal cell
Mesenchyme
Nuclei (cytology)
Nucleus pulposus
Pain
Proteoglycans
Stromal cells
Transforming growth factor-b1
Wnt
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Summary:Background: Multiple studies have examined the potential of growth factors (GF) to enable mesenchymal stromal cells (MSC) to nucleus pulposus (NP) cell-like cell differentiation. Here we screened a wide range of GF and GF combinations for supporting NP cell-like cell differentiation. Methods: Human MSC were stimulated using 86 different GF combinations of TGF-β1, -2, -3, GDF5, -6, Wnt3a, -5a, -11, and Shh. Differentiation potency was assessed by alcian blue assay and NP cell marker expression (e.g., COL2A1, CD24, etc.). The top four combinations and GDF5/TGF-β1 were further analyzed in 3D pellet cultures, on their ability to similarly induce NP cell differentiation. Results: Almost all 86 GF combinations showed their ability to enhance proteoglycan production in alcian blue assay. Subsequent qPCR analysis revealed TGF-β2/Wnt3a, TGF-β1/Wnt3a, TGF-β1/Wnt3a/GDF6, and Wnt3a/GDF6 as the most potent combinations. Although in pellet cultures, these combinations supported NP marker expression, none showed the ability to significantly induce chondrogenic NP matrix production. Only GDF5/TGF-β1 resulted in chondrogenic pellets with significantly enhanced glycosaminoglycan content. Conclusion: GDF5/TGF-β1 was suggested as an optimal GF combination for MSC to NP cell induction, although further assessment using a 3D and in vivo environment is required. Wnt3a proved promising for monolayer-based NP cell differentiation, although further validation is required.
ISSN:2076-3417
2076-3417
DOI:10.3390/app11083673