Abstract
Three-dimensional (3D) cell cultures combining multipotent mesenchymal stromal cells (MSC), tendon extracellular matrix scaffolds, and mechanical stimulation by a bioreactor have been used to induce tenogenic differentiation in vitro. Yet, these conditions alone do not mimic the environment of acute inflammatory tendon disease adequately, thus the results of such studies are not representatives for tendon regeneration after acute injury. In this chapter, we describe two different approaches to introduce inflammatory stimuli, comprising co-culture with leukocytes and supplementation with the cytokines IL-1 β and TNF-α. The presented in vitro model of inflammatory tendon disease could be used to study musculoskeletal pathophysiology and regeneration in more depth.
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References
Lui PPY (2015) Stem cell technology for tendon regeneration: current status, challenges, and future research directions. Stem Cells Cloning 8:163–174. https://doi.org/10.2147/SCCAA.S60832
Manning CN, Martel C, Sakiyama-Elbert SE et al (2015) Adipose-derived mesenchymal stromal cells modulate tendon fibroblast responses to macrophage-induced inflammation in vitro. Stem Cell Res Ther 6:74. https://doi.org/10.1186/s13287-015-0059-4
Burk J, Erbe I, Berner D et al (2014) Freeze-thaw cycles enhance decellularization of large tendons. Tissue Eng Part C Methods 20(4):276–284. https://doi.org/10.1089/ten.TEC.2012.0760
Cheng CW, Solorio LD, Alsberg E (2014) Decellularized tissue and cell-derived extracellular matrices as scaffolds for orthopaedic tissue engineering. Biotechnol Adv 32(2):462–484. https://doi.org/10.1016/j.biotechadv.2013.12.012
Youngstrom DW, Barrett JG (2016) Tendon differentiation on decellularized extracellular matrix under cyclic loading. Methods Mol Biol 1502:195–202. https://doi.org/10.1007/7651_2016_332
Roth SP, Glauche SM, Plenge A et al (2017) Automated freeze-thaw cycles for decellularization of tendon tissue—a pilot study. BMC Biotechnol 17(1):13. https://doi.org/10.1186/s12896-017-0329-6
Roth SP, Erbe I, Burk J (2018) Decellularization of large tendon specimens: combination of manually performed freeze-thaw cycles and detergent treatment. Methods Mol Biol 1577:227–237. https://doi.org/10.1007/7651_2017_49
Youngstrom DW, Rajpar I, Kaplan DL et al (2015) A bioreactor system for in vitro tendon differentiation and tendon tissue engineering. J Orthop Res 33(6):911–918. https://doi.org/10.1002/jor.22848
Burk J, Plenge A, Brehm W et al (2016) Induction of Tenogenic differentiation mediated by extracellular tendon matrix and short-term cyclic stretching. Stem Cells Int 2016:7342379. https://doi.org/10.1155/2016/7342379
Han P, Cui Q, Yang S et al (2017) Tumor necrosis factor-α and transforming growth factor-β1 facilitate differentiation and proliferation of tendon-derived stem cells in vitro. Biotechnol Lett 39(5):711–719. https://doi.org/10.1007/s10529-017-2296-3
Zhang K, Asai S, Yu B et al (2015) IL-1β irreversibly inhibits tenogenic differentiation and alters metabolism in injured tendon-derived progenitor cells in vitro. Biochem Biophys Res Commun 463(4):667–672. https://doi.org/10.1016/j.bbrc.2015.05.122
Brandt L, Schubert S, Scheibe P et al (2018) Tenogenic properties of Mesenchymal progenitor cells are compromised in an inflammatory environment. Int J Mol Sci 19(9). https://doi.org/10.3390/ijms19092549
Hillmann A, Paebst F, Brehm W et al (2019) A novel direct co-culture assay analyzed by multicolor flow cytometry reveals context- and cell type-specific immunomodulatory effects of equine mesenchymal stromal cells. PLoS One 14(6):e0218949. https://doi.org/10.1371/journal.pone.0218949
Cassano JM, Schnabel LV, Goodale MB et al (2018) Inflammatory licensed equine MSCs are chondroprotective and exhibit enhanced immunomodulation in an inflammatory environment. Stem Cell Res Ther 9(1):82. https://doi.org/10.1186/s13287-018-0840-2
Schulze-Tanzil G, Al-Sadi O, Wiegand E et al (2011) The role of pro-inflammatory and immunoregulatory cytokines in tendon healing and rupture: new insights. Scand J Med Sci Sports 21(3):337–351. https://doi.org/10.1111/j.1600-0838.2010.01265.x
Burk J, Ribitsch I, Gittel C et al (2013) Growth and differentiation characteristics of equine mesenchymal stromal cells derived from different sources. Vet J 195(1):98–106. https://doi.org/10.1016/j.tvjl.2012.06.004
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Schubert, S., Brandt, L., Burk, J. (2021). A 3D Dynamic In Vitro Model of Inflammatory Tendon Disease. In: Stock, P., Christ, B. (eds) In Vitro Models for Stem Cell Therapy. Methods in Molecular Biology, vol 2269. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1225-5_12
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DOI: https://doi.org/10.1007/978-1-0716-1225-5_12
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