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.

细胞外基质调节细胞存活、增殖和分化。体外二维细胞实验通常在塑料板或单一细胞外基质成分（如胶原蛋白或磷酸钙）的基底上进行。由于这些方法不包括细胞外基质蛋白或生长因子，它们无法模拟复杂的细胞微环境。细胞衍生的基质是一种替代平台，可以更好地代表体外的体内微环境。细胞衍生基质的标准去细胞化是通过结合化学和物理方法实现的。在这项研究中，我们比较了几种方法的脱细胞效果：氢氧化铵、十二烷基硫酸钠 (SDS) 或 Triton X-100 在 Saos-2 细胞基质上进行冷处理或热处理。我们发现含有 SDS 的方案在再细胞化过程中具有细胞毒性。47°C 的热处理没有细胞毒性，去除细胞成分，灭活碱性磷酸酶活性，并保持钙沉积水平。随后，我们研究了直接骨共培养系统在已建立的脱细胞 Saos-2 基质、磷酸钙无机基质和作为对照的塑料板中的分化效率。我们发现通过47℃热处理获得的脱细胞Saos-2细胞基质比无机基质和对照更好地增强破骨细胞分化和基质矿化。这种简单且低成本的方法使我们能够创建 Saos-2 脱细胞基质，可用作骨细胞生长和分化的体内样支持。