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首页> 外文期刊>Biomaterials >Stable immobilization of rat hepatocyte spheroids on galactosylated nanofiber scaffold.
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Stable immobilization of rat hepatocyte spheroids on galactosylated nanofiber scaffold.

机译：将大鼠肝细胞球体稳定固定在半乳糖基化纳米纤维支架上。

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Primary rat hepatocytes self-assemble into multi-cellular spheroids and maintain differentiated functions when cultured on a two-dimensional (2-D) substrate conjugated with galactose ligand. The aim of this study is to investigate how a functional nanofiber scaffold with surface-galactose ligand influences the attachment, spheroid formation and functional maintenance of rat hepatocytes in culture, as compared with the functional 2-D substrate. Highly porous nanofiber scaffolds comprising of fibers with an average diameter of 760nm were prepared by electrospinning of poly(epsilon-caprolactone-co-ethyl ethylene phosphate) (PCLEEP), a novel biodegradable copolymer. Galactose ligand with a density of 66nmol/cm(2) was achieved on the nanofiber scaffold via covalent conjugation to a poly(acrylic acid) spacer UV-grafted onto the fiber surface. Hepatocytes cultured on the galactosylated PCLEEP nanofiber scaffold exhibited similar functional profiles in terms of cell attachment, ammonia metabolism, albumin secretion and cytochrome P450 enzymatic activity as those on the functional 2-D substrate, although their morphologies are different. Hepatocytes cultured on galactosylated PCLEEP film formed 50-300mum spheroids that easily detached from surface upon agitation, whereas hepatocytes cultured on galactosylated nanofiber scaffold formed smaller aggregates of 20-100mum that engulfed the functional nanofibers, resulting in an integrated spheroid-nanofiber construct.
机译：在与半乳糖配体缀合的二维（2-D）底物上培养时，原代大鼠肝细胞自组装成多细胞球体，并维持分化的功能。这项研究的目的是研究与功能性2维底物相比，具有表面半乳糖配体的功能性纳米纤维支架如何影响培养中大鼠肝细胞的附着，球体形成和功能性维持。通过电纺丝一种新型的可生物降解的共聚物聚（ε-己内酯-乙基乙基磷酸乙烯酯）（PCLEEP），制备了平均孔径为760nm的高多孔纳米纤维支架。在纳米纤维支架上，通过与紫外线接枝到纤维表面的聚（丙烯酸）间隔基共价结合，在密度为66nmol / cm（2）的半乳糖上实现了配体。在半乳糖基化PCLEEP纳米纤维支架上培养的肝细胞在细胞附着，氨代谢，白蛋白分泌和细胞色素P450酶活性方面表现出与功能性2-D底物相似的功能谱，尽管它们的形态不同。在半乳糖基化PCLEEP膜上培养的肝细胞形成50-300μm的球状体，这些球状体在搅拌时很容易从表面分离，而在半乳糖基化的纳米纤维支架上培养的肝细胞形成了20-100μm的较小聚集体，吞噬了功能性纳米纤维，形成了一体化的球状-纳米纤维构建体。

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