Laboratory of Stem Cell Research and Application, University of Science, VNUHCM, Ho Chi Minh City, Vietnam.
Vietnam National University, Ho Chi Minh City, Vietnam.
Am J Physiol Cell Physiol. 2023 Jun 1;324(6):C1213-C1222. doi: 10.1152/ajpcell.00001.2023. Epub 2023 Apr 17.
Scaffold-based culture is necessary for hepatic stellate cells (HSCs) because HSCs are promptly autoactivated under plastic conditions. Our research aims to investigate the potential and role of fibrin scaffold in reducing autoactivation, maintaining cell function, and extending the in vitro culture time of primary HSCs. HSCs were isolated from BALB/c mice and cultured on the surface of plastic, Matrigel, and fibrin gel. HSC's characteristics, including recovery, morphology, proliferation, lipid droplet (LD) storage, and activation were evaluated. Cell recovery was 86%, 80%, and 60% in fibrin, Matrigel, and plastic, respectively ( < 0.05). HSCs cultured on a plastic dish were autoactivated until with high proliferation, loss of cytoplasmic LD lipid droplets, and increased expression of activation markers, including alpha-smooth muscle actin (α-sma) and collagen type I. In contrast, these phenomena were reduced in Matrigel and fibrin-based cultures ( < 0.05). HSC culture in fibrin scaffold was associated with altered expression of cell adhesion molecules, including increased and inhibited . HSCs were more stellate-like in morphology in fibrin than in the Matrigel scaffold. Interestingly, fibrin-scaffold-embedded culture was able to maintain HSC quiescent state for up to 14 days in vitro. Fibrin gel could provide a potential scaffold for primary HSC culture while preserving cell function and extending primary HSC in vitro culture time. Fibrin gel is appropriate for maintaining quiescence characteristics in primary culture of mouse hepatic stellate cells. Embedded culture of hepatic stellate cells in fibrin gel simulates in vivo cell morphology. Stiffness and adhesion molecules of fibrin gel play a crucial role in the hepatic stellate cell's primary culture.
支架培养是肝星状细胞（HSCs）所必需的，因为 HSCs 在塑料条件下会迅速自动激活。我们的研究旨在探讨纤维蛋白支架在减少自动激活、维持细胞功能和延长原代 HSCs 体外培养时间方面的潜力和作用。我们从 BALB/c 小鼠中分离 HSCs，并在塑料、Matrigel 和纤维蛋白凝胶表面培养。评估了 HSCs 的特性，包括恢复、形态、增殖、脂滴（LD）储存和激活。纤维蛋白、Matrigel 和塑料中 HSC 的回收率分别为 86%、80%和 60%（<0.05）。在塑料培养皿中培养的 HSCs 会自动激活，直到[时间]，表现为高增殖、细胞质 LD 脂质滴丢失和激活标志物（包括α-平滑肌肌动蛋白（α-sma）和 I 型胶原）表达增加。相比之下，这些现象在 Matrigel 和纤维蛋白基培养物中减少（<0.05）。纤维蛋白支架培养的 HSC 与细胞黏附分子的表达改变有关，包括增加[分子名称]和抑制[分子名称]。纤维蛋白中的 HSCs 形态更具星状，比 Matrigel 支架中的更明显。有趣的是，纤维蛋白支架包埋培养可使 HSCs 在体外保持静止状态长达 14 天。纤维蛋白凝胶可为原代 HSC 培养提供潜在的支架，同时保持细胞功能并延长原代 HSC 的体外培养时间。纤维蛋白凝胶适合维持原代培养的小鼠肝星状细胞的静止特征。纤维蛋白凝胶中肝星状细胞的包埋培养模拟了体内细胞形态。纤维蛋白凝胶的硬度和黏附分子在肝星状细胞的原代培养中起着关键作用。
