Charité Universitätsmedizin Berlin, Berlin-Brandenburg Center für Regenerative Therapies, Augustenburger Platz 1, 13353, Berlin, Germany.
King Faisal Specialist Hospital and Research Centre, Comparative Medicine Department, Zahrawi Street, Riyadh, 11211, Saudi Arabia.
Biomaterials. 2019 Sep;216:119283. doi: 10.1016/j.biomaterials.2019.119283. Epub 2019 Jun 18.
Extracellular matrix (ECM) provides a scaffold for cells and tissues, but also supports organogenesis and tissue remodeling. The required instructive properties of the ECM to interact with cells depend on matrix architecture, structural proteins and functional matrix components such as growth factors, providing spatial, chemical and functional cues. Decellularized ECM (dECM) has been proposed as an instructive material that promotes tissue regeneration. We investigated the instructive ECM elements preserved in dECM and necessary to promote endothelial differentiation of human induced pluripotent stem cells (hiPSC). We show that detergent-decellularized human kidney ECM remains structurally intact and carries a number of heparin-binding growth factors, including FGF2, VEGF, BMP2, HGF, EGF, PDGF-BB and TGFβ, albeit at reduced levels compared to native tissues. Clearance of these heparin-binding factors, or heparan-sulfate proteoclycans from ECM resulted in massively reduced differentiation of hiPSC, suggesting that remaining structural dECM proteins such as laminin, collagen or fibronectin alone are not instructive. In contrast, replenishing dECM with VEGF replaced medium-supplemented VEGF and resulted in more efficient differentiation of hiPSC into endothelial cells, and even in the absence of other culture-supplemented differentiation factors dECM alone was superior to geltrex. In conclusion, conditioning of dECM with specific growth factors acting as functional cues may allow to generate functional niches by selective promotion of cell attachment, survival and differentiation.
细胞外基质 (ECM) 为细胞和组织提供支架，但也支持器官发生和组织重塑。ECM 与细胞相互作用所需的指令特性取决于基质结构、结构蛋白和功能性基质成分，如生长因子，提供空间、化学和功能线索。去细胞细胞外基质 (dECM) 已被提议作为一种促进组织再生的指令性材料。我们研究了保留在 dECM 中并促进人诱导多能干细胞 (hiPSC) 内皮分化所需的指令性 ECM 元素。我们表明，去污剂去细胞化的人肾 ECM 仍然保持结构完整，并携带多种肝素结合生长因子，包括 FGF2、VEGF、BMP2、HGF、EGF、PDGF-BB 和 TGFβ，尽管与天然组织相比水平降低。这些肝素结合因子或 ECM 中的硫酸乙酰肝素蛋白聚糖的清除导致 hiPSC 的分化大大减少，这表明剩余的结构 dECM 蛋白，如层粘连蛋白、胶原蛋白或纤维连接蛋白本身没有指令性。相比之下，用 VEGF 补充 dECM 取代了培养基补充的 VEGF，从而更有效地将 hiPSC 分化为内皮细胞，即使在没有其他培养补充分化因子的情况下，dECM 本身也优于 geltrex。总之，用作为功能线索的特定生长因子对 dECM 进行调理，可能允许通过选择性促进细胞附着、存活和分化来产生功能性龛。
