Evaluating the Effects of Dithiothreitol and Fructose on Cell Viability and Function of Cryopreserved Primary Rat Hepatocytes and HepG2 Cell Line.
Evaluating the Effects of Dithiothreitol and Fructose on Cell Viability and Function of Cryopreserved Primary Rat Hepatocytes and HepG2 Cell Line.
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DOI：
10.5812/hepatmon.7824
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发表时间：
2013-01
影响因子：
0.6
通讯作者：
Azarpira N
中科院分区：
文献类型：
--
作者：
Aghdai MH;Jamshidzadeh A;Nematizadeh M;Behzadiannia M;Niknahad H;Amirghofran Z;Esfandiari E;Azarpira N
肝细胞用作体外模型以评价药物代谢。人肝细胞移植已被认为是急性肝功能衰竭的临时治疗方法。优化冷冻方法对于保存细胞活力和功能至关重要，而细胞活力和功能通常是通过冷冻保存来实现的。本研究旨在探讨DTT和果糖对原代大鼠肝细胞和HepG2细胞的冷冻保护作用。将新鲜大鼠肝细胞和HepG2细胞系分别与果糖（100和200 mM）和二硫苏糖醇（DTT）（25、50、100、250和500 μM）在37 ℃下孵育1小时和3小时。将预孵育的肝细胞冷冻保存两周。解冻后测定肝细胞活力和功能，并将结果与对照组进行比较。用200 mM果糖预孵育1 h后，大鼠肝细胞和HepG2细胞的存活率均显著增加。250 μM和500 μM）在两种细胞类型中均提高了解冻后的活力和功能（P < 0.001）。在大鼠肝细胞中，与果糖或DTT预孵育后，未观察到白蛋白、尿素产生和LDH泄漏的显著变化。在HepG2细胞中，用DTT预孵育（500 μM，1小时）后，白蛋白和尿素产量显著增加。DTT（250和500 μM，1小时）组大鼠肝细胞和HepG2细胞中的GSH含量显著增加。在冻存前用果糖和DTT孵育肝细胞可以增加解冻后的细胞活力和功能。
Hepatocytes are used as an in vitro model to evaluate drug metabolism. Human hepatocyte transplant has been considered as the temporary treatment of acute liver failure. Optimization freezing methods is very important to preserve both cell viability and function which are achieved by cryopreservation mostly always. The present study aimed to investigate the cryoprotective effect of DTT and fructose on primary rat hepatocytes and HepG2 cells. Both fresh rat hepatocytes and HepG2 cell line were incubated with fructose (100 and 200 mM) and dithiothreitol (DTT) (25, 50, 100, 250, and 500 μM) at 37°C for 1 and 3 hours, respectively. The preincubated hepatocytes were cryopreserved for two weeks. Hepatocytes viability and function were determined post thawing and the results were compared with the control group. The viability of both rat hepatocytes and HepG2 cells were significantly increased after one hour preincubation with fructose 200 mM. Preincubation with DTT (50 μM, 100 μM. 250 μM and 500 μM) improved the viability and function upon thawing in both cell types (P < 0.001). In rat hepatocytes, no significant change was observed in albumin, urea production, and LDH leakage after preincubation with fructose or DTT. In HepG2 cells, albumin and urea production were significantly increased after preincubation with DTT (500 μM, 1 hour). The GSH content was significantly increased in DTT (250 and 500 μM, 1 hour) groups in both rat hepatocyte and HepG2 cells. Incubation of hepatocytes with fructose and DTT prior to the cryopreservation can increase the cell viability and function after thawing.
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影响因子：
3.3
作者：
ADAMS, RM;WANG, M;LEDLEY, FD
通讯作者：
LEDLEY, FD
影响因子：
3.3
作者：
Fujita, R;Hui, T;Demetriou, AA
通讯作者：
Demetriou, AA
影响因子：
4.8
作者：
Gauthaman, Kalamegam;Fong, Chui-Yee;Bongso, Ariff
通讯作者：
Bongso, Ariff
影响因子：
5.1
作者：
Guillouzo, A;Rialland, L;Guyomard, C
通讯作者：
Guyomard, C
影响因子：
4.8
作者：
Biroccio, A;Benassi, B;Zupi, G
通讯作者：
Zupi, G