Three-dimensional co-culture of rat hepatocyte spheroids and NIH/3T3 fibroblasts enhances hepatocyte functional maintenance
Three-dimensional co-culture of rat hepatocyte spheroids and NIH/3T3 fibroblasts enhances hepatocyte functional maintenance
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DOI：
10.1016/j.actbio.2005.04.003
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发表时间：
2005-07-01
影响因子：
9.7
通讯作者：
Mao, HQ
中科院分区：
文献类型：
--
作者：
Lu, HF;Chua, KN;Mao, HQ
原代肝细胞在培养中的功能维持可以通过几种不同的方法来改善，包括优化细胞外基质微环境、介质组成和细胞-细胞相互作用，包括同型和异型。利用半乳糖修饰的表面，我们开发了一种将这两种方法结合起来的方法，通过将大鼠原代肝细胞球体与NIH/3T3小鼠成纤维细胞共同培养。将肝细胞在半乳糖化聚偏氟乙烯膜上培养3d，制成球形，然后接种NIH/3T3细胞并与其共培养。结果表明，尽管NIH/3T3细胞单独对半乳糖修饰的PVDF表面反应较差，并表现出有限的附着，但NIH/3T3成纤维细胞附着在肝细胞球体的周围并在其周围增殖。与单独培养的肝细胞球体相比，共培养的肝细胞球体表现出更高的肝脏特异性功能。在无血清培养(第3天)和有血清培养(第4天)中，该共培养体系的白蛋白分泌水平在第11天达到峰值，分别是球形同源培养对照组的1.8倍和2.9倍。白蛋白的分泌功能至少维持了两周，是球形同源培养油第13天的5.1倍(无血清培养)和17.8倍(有血清培养)。同样，共培养体系在无血清培养和有血清培养的第14天表达的细胞色素P450酶活性分别是同种培养对照的5.5倍和3.1倍。总之，这种独特的共培养体系显示了同型细胞-细胞相互作用、异型细胞-细胞相互作用、细胞-底物相互作用和可溶性刺激在肝细胞功能维持中的协同作用。(C)2005年Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Functional maintenance of primary hepatocytes in culture can be improved by several distinct approaches involving optimization of the extracellular matrix microenvironment, media composition and cell-cell interactions, both homotypic and heterotypic. Using a galactose-decorated surface, we have developed a method to combine these two approaches by co-culturing rat primary hepatocyte spheroids with NIH/3T3 mouse fibroblast cells. Spheroids were performed by culturing hepatocytes for 3 days on galactosylated poly(vinylidene difluoride) membrane; NIH/3T3 cells were subsequently seeded and co-cultured with the spheroids. Results showed that although NIH/3T3 cells alone responded poorly to the galactosylated PVDF surface and displayed limited attachment, NIH/3T3 fibroblasts attached to the periphery of the hepatocyte spheroids and proliferated around them. Co-cultured hepatocyte spheroids exhibited significantly higher liver-specific functions as compared to spheroids cultured alone. Albumin secretion level in this co-culture system peaked oil day 11, which was 1.8- and 2.9-times higher than the peak expression level in spheroid homo-culture control in serum-free (day 3) and serum-containing media (day 4), respectively. The albumin secretion function was maintained for at least two weeks; it was 5.1 (in serum-free medium) and 17.8 (in serum-containing medium) times higher than spheroid homo-culture oil day 13. Similarly, the co-culture system also expressed approximately 5.5- and 3.1-times higher 3-methylcholanthrene-induced cytochrome P450 enzymatic activity oil day 14 as compared to the homo-culture control in serum-free and serum-containing medium, respectively. In conclusion, this unique co-culture system demonstrated the synergistic roles of homotypic cell-cell interaction, heterotypic cell-cell interaction, cell-substrate interaction and soluble stimuli in hepatocyte functional maintenance. (c) 2005 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.