Gregory Carl A, McNeill Eoin P, Pan Simin
Department of Molecular and Cellular Medicine, Institute for Regenerative Medicine, Texas A&M Health Science Center, College Station, TX, United States.
Department of Molecular and Cellular Medicine, Institute for Regenerative Medicine, Texas A&M Health Science Center, College Station, TX, United States.
Methods Cell Biol. 2020;156:15-43. doi: 10.1016/bs.mcb.2019.10.009. Epub 2019 Dec 10.
Bone is a composite material consisting primarily of cells, extracellular matrices, accessory proteins and the complex calcium phosphate salt hydroxyapatite. Collectively, the extracellular network of proteins and accessory molecules that provide the organic component of bone tissue is referred to as the osteogenic extracellular matrix (OECM). OECM provides tensile strength and increases the durability of bone, but the OECM also serves as an attachment site and regulatory substrate for cells and a repository for growth factors and cytokines. Increasingly, purified OECM generated by osteogenic cells in culture has attracted interest because it has the capacity to improve the growth and viability of attached cells, enhances the osteogenic program in vitro and in vivo, and shows great promise as a therapeutic tool for orthopedic tissue engineering. This chapter will describe fundamental protocols for the selection and culture of osteogenic cells and conditions for their osteogenic differentiation, and the synthesis, purification and characterization of OECM. Some examples of immobilization to surfaces for the purpose of two- and three-dimensional culture will also be described.
骨是一种复合材料，主要由细胞、细胞外基质、辅助蛋白和复杂的磷酸钙盐羟基磷灰石组成。总体而言，提供骨组织有机成分的蛋白质和辅助分子的细胞外网络被称为成骨细胞外基质（OECM）。OECM提供抗张强度并增加骨的耐久性，但OECM还作为细胞的附着位点和调节底物以及生长因子和细胞因子的储存库。越来越多的由培养的成骨细胞产生的纯化OECM引起了人们的兴趣，因为它有能力改善附着细胞的生长和活力，在体外和体内增强成骨程序，并作为骨科组织工程的治疗工具显示出巨大的前景。本章将描述成骨细胞的选择和培养的基本方案及其成骨分化的条件，以及OECM的合成、纯化和表征。还将描述用于二维和三维培养目的固定到表面的一些示例。
