Department of Environmental Sciences and Engineering, University of North Carolina at Chapel Hill, NC 27599, USA.
Toxicol Appl Pharmacol. 2010 Dec 15;249(3):208-16. doi: 10.1016/j.taap.2010.09.012. Epub 2010 Sep 24.
Numerous studies support the fact that a genetically diverse mouse population may be useful as an animal model to understand and predict toxicity in humans. We hypothesized that cultures of hepatocytes obtained from a large panel of inbred mouse strains can produce data indicative of inter-individual differences in in vivo responses to hepato-toxicants. In order to test this hypothesis and establish whether in vitro studies using cultured hepatocytes from genetically distinct mouse strains are feasible, we aimed to determine whether viable cells may be isolated from different mouse inbred strains, evaluate the reproducibility of cell yield, viability and functionality over subsequent isolations, and assess the utility of the model for toxicity screening. Hepatocytes were isolated from 15 strains of mice (A/J, B6C3F1, BALB/cJ, C3H/HeJ, C57BL/6J, CAST/EiJ, DBA/2J, FVB/NJ, BALB/cByJ, AKR/J, MRL/MpJ, NOD/LtJ, NZW/LacJ, PWD/PhJ and WSB/EiJ males) and cultured for up to 7 days in traditional 2-dimensional culture. Cells from B6C3F1, C57BL/6J, and NOD/LtJ strains were treated with acetaminophen, WY-14,643 or rifampin and concentration-response effects on viability and function were established. Our data suggest that high yield and viability can be achieved across a panel of strains. Cell function and expression of key liver-specific genes of hepatocytes isolated from different strains and cultured under standardized conditions are comparable. Strain-specific responses to toxicant exposure have been observed in cultured hepatocytes and these experiments open new opportunities for further developments of in vitro models of hepatotoxicity in a genetically diverse population.
大量研究支持这样一个事实，即遗传多样性的小鼠群体可作为一种动物模型，有助于理解和预测人类的毒性。我们假设，从大量近交系小鼠品系中获得的肝细胞培养物可以产生数据，表明对肝毒性物质的体内反应存在个体间差异。为了验证这一假设并确定使用遗传上不同的小鼠品系培养的肝细胞进行体外研究是否可行，我们旨在确定是否可以从不同的近交系小鼠中分离出有活力的细胞，评估细胞产量、活力和功能在随后的分离中的可重复性，并评估该模型用于毒性筛选的实用性。我们从 15 个品系的小鼠（A/J、B6C3F1、BALB/cJ、C3H/HeJ、C57BL/6J、CAST/EiJ、DBA/2J、FVB/NJ、BALB/cByJ、AKR/J、MRL/MpJ、NOD/LtJ、NZW/LacJ、PWD/PhJ 和 WSB/EiJ 雄性）中分离出肝细胞，并在传统的二维培养中培养长达 7 天。用对乙酰氨基酚、WY-14,643 或利福平处理 B6C3F1、C57BL/6J 和 NOD/LtJ 品系的细胞，建立对活力和功能的浓度反应。我们的数据表明，在一组品系中可以实现高产量和高活力。在标准化条件下从不同品系分离并培养的肝细胞的功能和关键肝脏特异性基因的表达是可比的。在培养的肝细胞中观察到对毒物暴露的品系特异性反应，这些实验为在遗传多样性人群中进一步开发体外肝毒性模型开辟了新的机会。
