A simple method for decellularizing a cell-derived matrix for bone cell cultivation and differentiation

Autor: Weidong Weng, Filippo Zanetti, David Bovard, Bianca Braun, Sabrina Ehnert, Tatiana Uynuk-Ool, Tina Histing, Julia Hoeng, Andreas K. Nussler, Romina H. Aspera-Werz
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Journal of Materials Science: Materials in Medicine, Vol 32, Iss 9, Pp 1-13 (2021)
Druh dokumentu: article
ISSN: 0957-4530
1573-4838
DOI: 10.1007/s10856-021-06601-y
Popis: Abstract The extracellular matrix regulates cell survival, proliferation, and differentiation. In vitro two-dimensional cell experiments are typically performed on a plastic plate or a substrate of a single extracellular matrix constituent such as collagen or calcium phosphate. As these approaches do not include extracellular matrix proteins or growth factors, they fail to mimic a complex cell microenvironment. The cell-derived matrix is an alternative platform for better representing the in vivo microenvironment in vitro. Standard decellularization of a cell-derived matrix is achieved by combining chemical and physical methods. In this study, we compared the decellularization efficacy of several methods: ammonium hydroxide, sodium dodecyl sulfate (SDS), or Triton X-100 with cold or heat treatment on a matrix of Saos-2 cells. We found that the protocols containing SDS were cytotoxic during recellularization. Heat treatment at 47 °C was not cytotoxic, removed cellular constituents, inactivated alkaline phosphatase activity, and maintained the levels of calcium deposition. Subsequently, we investigated the differentiation efficiency of a direct bone coculture system in the established decellularized Saos-2 matrix, an inorganic matrix of calcium phosphate, and a plastic plate as a control. We found that the decellularized Saos-2 cell matrix obtained by heat treatment at 47 °C enhanced osteoclast differentiation and matrix mineralization better than the inorganic matrix and the control. This simple and low-cost method allows us to create a Saos-2 decellularized matrix that can be used as an in vivo-like support for the growth and differentiation of bone cells.
Databáze: Directory of Open Access Journals
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