Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function.
Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function.
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DOI：
10.1002/hep.22674
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发表时间：
2009-02
期刊：
影响因子：
13.5
通讯作者：
Nyberg, Scott L.
中科院分区：
文献类型：
--
作者：
Brophy, Colleen M.;Luebke-Wheeler, Jennifer L.;Amiot, Bruce P.;Khan, Harris;Remmel, Rory P.;Rinaldo, Piero;Nyberg, Scott L.
原代肝细胞球形培养为体外肝脏研究创造了一种有效的三维组织结构。球状体具有结构极性和功能性胆管，分化功能正常。因此，肝细胞球体已被提出作为多种诊断、发现和治疗应用的细胞来源，如生物人工肝。采用一种新的摇摆技术诱导球体形成，在培养的14天内检测了大鼠肝细胞球体形成动力学、细胞-细胞粘附、基因表达和生化活性。有证据表明，与传统的旋转系统相比，摇摆悬浮培养中球体的形成速度更快，非粘附性肝细胞也更少。摇培养的肝球细胞稳定表达了白蛋白合成、尿素循环、I、II期代谢酶和凝血因子等242种肝脏相关基因的80%以上。摇球形肝细胞对组织培养塑料和胶原蛋白的生化活性优于单层肝细胞。综上所述，用摇臂技术形成球体比旋转技术更快、更有效。摇臂形成的球体似乎适合应用于生物人工肝脏或作为体外肝组织结构。
The culture of primary hepatocytes as spheroids creates an efficient 3-dimensional tissue construct for hepatic studies in vitro. Spheroids possess structural polarity and functional bile canaliculi with normal differentiated function. Thus, hepatocyte spheroids have been proposed as the cell source in a variety of diagnostic, discovery, and therapeutic applications, such as a bioartificial liver. Using a novel rocking technique to induce spheroid formation, kinetics of spheroid formation, cell-cell adhesion, gene expression and biochemical activities of rat hepatocyte spheroids were tested over 14 days of culture. Evidence was provided that the formation of spheroids occurred faster and with fewer non-adherent hepatocytes in rocked suspension culture compared to a traditional rotational system. Hepatocyte spheroids in rocked culture showed stable expression of over 80% of 242 liver-related genes including those of albumin synthesis, urea cycle, phase I and II metabolic enzymes, and clotting factors. Biochemical activity of rocked spheroid hepatocytes was superior to monolayer culture of hepatocytes on tissue culture plastic and collagen. In conclusion, spheroid formation by rocker technique was more rapid and more efficient than rotational technique. Rocker formed spheroids appear suitable for application in a bioartificial liver or as an in vitro liver tissue construct.
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影响因子：
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