University of Groningen, University Medical Center Groningen, Department of Gastroenterology and Hepatology, Groningen, the Netherlands; University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, the Netherlands.
University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, the Netherlands; University of Groningen, University Medical Centre Groningen, W.J. Kolff Institute for Biomedical Engineering and Materials Science, Groningen, the Netherlands; University of Groningen, University Medical Centre Groningen, Department of Biomedical Engineering, Groningen, the Netherlands.
Int J Biol Macromol. 2024 Oct;278(Pt 2):134717. doi: 10.1016/j.ijbiomac.2024.134717. Epub 2024 Aug 12.
Liver sinusoidal endothelial cells (LSECs) are key targets for addressing metabolic dysfunction-associated steatotic liver disease (MASLD). However, isolating and culturing primary LSECs is challenging due to rapid dedifferentiation, resulting in loss of function. The extracellular matrix (ECM) likely plays a crucial role in maintaining the fate and function of LSECs. In this study, we explored the influence of liver-ECM (L-ECM) on liver cells and developed culture conditions that maintain the differentiated function of liver cells in vitro for prolonged periods. Porcine liver-derived L-ECM, containing 34.9 % protein, 0.045 % glycosaminoglycans, and negligible residual DNA (41.2 ng/mg), was utilized to culture primary rat liver cells in generated hydrogels. Proteomic analyses and molecular weight distribution of proteins of solubilized L-ECM revealed the typical diverse ECM core matrisome, with abundant collagens. L-ECM hydrogels showed suitable stiffness and stress relaxation properties. Furthermore, we demonstrated that collagen-rich L-ECM hydrogels enhanced LSECs' and hepatocytes' viability, and reduced the dedifferentiation rate of LSECs. In addition, hepatocyte function was maintained longer by culture on L-ECM hydrogels compared to traditional culturing. These beneficial effects are likely attributed to the bioactive macromolecules including collagens, and mechanical and microarchitectural properties of the L-ECM hydrogels.
肝窦内皮细胞（LSECs）是解决代谢功能障碍相关脂肪性肝病（MASLD）的关键靶点。然而，由于快速去分化，导致功能丧失，分离和培养原代 LSECs 具有挑战性。细胞外基质（ECM）可能在维持 LSECs 的命运和功能方面发挥着重要作用。在这项研究中，我们探讨了肝 ECM（L-ECM）对肝细 胞的影响，并开发了体外培养条件，以维持肝细 胞的分化功能，延长其体外培养时间。我们利用来源于猪肝脏的 L-ECM 培养原代大鼠肝细 胞，L-ECM 含有 34.9%的蛋白质、0.045%的糖胺聚糖和可忽略不计的残留 DNA（41.2ng/mg），并将其包被在生成的水凝胶中。L-ECM 可溶性成分的蛋白质组学分析和分子量分布揭示了典型的多样化 ECM 核心细胞外基质，其中富含胶原蛋白。L-ECM 水凝胶具有合适的硬度和应力松弛性能。此外，我们证明富含胶原蛋白的 L-ECM 水凝胶增强了 LSECs 和肝细胞的活力，并降低了 LSECs 的去分化率。此外，与传统培养相比，在 L-ECM 水凝胶上培养可延长肝细胞的功能维持时间。这些有益效果可能归因于包括胶原蛋白在内的生物活性大分子，以及 L-ECM 水凝胶的机械和微观结构特性。
