Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
Artif Organs. 2010 Jun;34(6):462-72. doi: 10.1111/j.1525-1594.2009.00885.x. Epub 2010 Apr 27.
Various metabolic tests were compared for the performance characterization of a liver cell bioreactor as a routine function assessment of cultures in a standby for patient application in clinical studies. Everyday quality assessment (QA) is essential to ensure a continuous level of cellular functional capacity in the development of hepatic progenitor cell expansion systems providing cells for regenerative medicine research; it is also of interest to meet safety requirements in bioartificial extracorporeal liver support systems under clinical evaluation. Quality criteria for the description of bioreactor cultures were developed using primary porcine liver cells as a model. Porcine liver cells isolated by collagenase perfusion with an average of 3 x 10(9) primary cells were used in 39 bioreactors for culture periods up to 33 days. Measurements of monoethylglycinexylidide synthesis and elimination of lidocaine, galactose elimination, and sorbitol elimination proved to be useful for routine QA of primary liver cell cultures. We demonstrate two methods for dispensing test substances, bolus administration and continuous, steady-state administration. Bolus test data were grouped in Standard, Therapy, Infection/Contamination, and Cell-free control groups. Statistical analyses show significant differences among all groups for every test substance. Post hoc comparisons indicated significant differences between Standard and Cell-free groups for all elimination parameters. For continuous tests, results were categorized according to number of culture days and time-dependent changes were analyzed. Continuous administration enables a better view of culture health and the time dependency of cellular function, whereas bolus administration is more flexible. Both procedures can be used to define cell function. Assessment of cellular function and bioreactor quality can contribute significantly to the quality of experimental or clinical studies in the field of hepatic bioreactor development.
各种代谢测试被比较,以表征作为患者临床研究应用备用的培养物的常规功能的肝细胞生物反应器。日常质量评估(QA)对于确保肝祖细胞扩增系统中细胞功能的连续水平是必要的,该系统为再生医学研究提供细胞;对于在临床评估下的生物人工体外肝脏支持系统满足安全性要求也很重要。使用原代猪肝细胞作为模型,开发了用于描述生物反应器培养物的质量标准。通过胶原酶灌注分离的猪肝细胞,平均每个生物反应器使用 3 x 10(9)个原代细胞,培养时间长达 33 天。测定单乙基甘氨酸二甲酰胺合成和利多卡因消除、半乳糖消除和山梨醇消除,证明对原代肝细胞培养物的常规 QA 有用。我们展示了两种分配测试物质的方法,即单次给药和连续稳态给药。单次给药测试数据分为标准、治疗、感染/污染和无细胞对照组。统计分析表明,每种测试物质在所有组之间均存在显著差异。事后比较表明,所有消除参数的标准组和无细胞组之间存在显著差异。对于连续测试,根据培养天数对结果进行分类,并分析时间依赖性变化。连续给药能够更好地观察培养物的健康状况和细胞功能的时间依赖性,而单次给药更灵活。这两种方法都可用于定义细胞功能。评估细胞功能和生物反应器质量可以为肝生物反应器开发领域的实验或临床研究的质量做出重大贡献。