Assessment of Hepatotoxic Potential of Cyanobacterial Toxins Using 3D In Vitro Model of Adult Human Liver Stem Cells
Assessment of Hepatotoxic Potential of Cyanobacterial Toxins Using 3D In Vitro Model of Adult Human Liver Stem Cells
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DOI：
10.1021/acs.est.8b02291
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发表时间：
2018-09-04
影响因子：
11.4
通讯作者：
Babica, Pavel
中科院分区：
文献类型：
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作者：
Basu, Amrita;Dydowiczova, Arleta;Babica, Pavel
蓝藻毒素微囊藻毒素-LR(MC-LR)和圆柱状精液蛋白(CyN)是一种危险的水环境污染物，对人体有很强的毒性。然而，体外单层培养的肝细胞系被发现概括了很差的主要肝细胞特异性功能，并且不能充分预测MC-LR和CyN的肝毒性效应。我们利用人端粒酶永生化的成人肝干细胞HL1-HT1的三维(3D)无支架球形培养来评估MC-LR和CyN的肝毒性潜力。在单层培养的HL-HT1细胞中，MC-LR不产生细胞毒作用(EC_(50)和10微摩尔/L)，而Cyn则抑制细胞的生长和存活(48小时-96小时的EC_(50)接近5.5-0.6微摩尔/L)。两种氰化毒素均抑制小球体的生长和存活，并与球体表面的气泡和解体有关。在大的成熟球体中也观察到肝球样损伤和活性降低，MC-LR的活性96h-EC50值为0.04micromol/L，Cyn的EC50值为0.1micromol/L，未观察到有效浓度
Cyanotoxins microcystin-LR (MC-LR) and cylindrospermopsin (CYN) represent hazardous waterborne contaminants and potent human hepatotoxins. However, in vitro monolayer cultures of hepatic cell lines were found to recapitulate, poorly, major hepatocyte-specific functions and inadequately predict hepatotoxic effects of MC-LR and CYN. We utilized 3-dimensional (3D), scaffold-free spheroid cultures of human telomerase-immortalized adult liver stem cells HL1-hT1 to evaluate hepatotoxic potential of MC-LR and CYN. In monolayer cultures of HL1-hT1 cells, MC-LR did not induce cytotoxic effects (EC50 > 10 micromol/L), while CYN inhibited cell growth and viability (48h-96h EC50 approximate to 5.5-0.6 micromol/L). Growth and viability of small growing spheroids were inhibited by both cyanotoxins (>= 0.1 micromol/L) and were associated with blebbing and disintegration at the spheroid surface. Hepatospheroid damage and viability reduction were observed also in large mature spheroids, with viability 96h-EC50 values being 0.04 micromol/L for MC-LR and 0.1 micromol/L for CYN, and No Observed Effect Concentrations