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首页> 外文期刊>Biomolecular engineering >The influence of medium composition and matrix on long-term cultivation of primary porcine and human hepatocytes.
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The influence of medium composition and matrix on long-term cultivation of primary porcine and human hepatocytes.

机译：培养基组成和基质对原代猪和人肝细胞长期培养的影响。

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摘要

The differentiated hepatocyte phenotype remains difficult to maintain in culture. The duration over which phenotypically stable hepatocytes can be cultured ranges from a couple of days to a few weeks. Shortcomings in medium formulation may be a factor in this lack of success. We have investigated effects of medium formulation on primary porcine and human hepatocyte cultures. We tested seven culture medium compositions (DMEM, ExCell 400, HepatoZYME-SFM, L-15 Leibovitz, SF-3, Waymouth, and Williams' E) and the effects of serum, fibronectin and biomatrix in a sandwich culture configuration. Albumin, urea, cholesterol, GOT, GPT, LDH and triglyceride concentrations were measured over 14 days. For both human and porcine cultures, the best results were obtained with SF-3 medium. Cells cultivated with Williams' E medium and FCS had good morphology and synthetic function during the first days of culture. However, continued addition of serum, was associated with a subsequent loss of differentiated phenotype. Addition of fibronectin was associated with improved function in cultures maintained in SF-3 medium whilst biomatrix had no effect. In contrast, addition of fibronectin did not influence cultures maintained in Williams' E medium, but cultures with biomatrix were associated with improved function at longer time points.
机译：分化的肝细胞表型仍然难以在培养中维持。表型稳定的肝细胞可以培养的持续时间为几天到几周。培养基配方的不足可能是缺乏成功的一个因素。我们已经研究了培养基配方对原代猪和人肝细胞培养的影响。我们测试了七种培养基成分（DMEM，ExCell 400，HepatoZYME-SFM，L-15 Leibovitz，SF-3，Waymouth和Williams'E）以及血清，纤连蛋白和生物基质在三明治培养结构中的作用。在14天内测量白蛋白，尿素，胆固醇，GOT，GPT，LDH和甘油三酸酯的浓度。对于人和猪培养物，使用SF-3培养基均可获得最佳结果。用威廉姆斯E培养基和FCS培养的细胞在培养的第一天具有良好的形态和合成功能。然而，继续添加血清与随后的分化表型丧失有关。纤连蛋白的添加与在SF-3培养基中维持的培养物中功能的改善有关，而生物基质没有作用。相反，添加纤连蛋白并不会影响威廉姆斯E培养基中维持的培养，但是具有生物基质的培养在较长时间点上与功能改善相关。

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