公开(公告)号：US06228595B1

公开(公告)日：2001-05-08

申请号：US09619444

申请日：2000-07-19

申请人： Dale Lynn Morris ,  Julio Cesar Davila

发明人： Dale Lynn Morris ,  Julio Cesar Davila

IPC分类号： C12Q168

CPC分类号： C12Q1/686 ,  C12Q1/6876 ,  C12Q1/6888 ,  C12Q2600/142 ,  C12Q2600/158 ,  C12Q2537/143 ,  C12Q2521/107

摘要： Hepatocyte culturing system, primer sets and an analytical method for selectively detecting and quantitatively assessing the levels of mRNA expression of the major isoenzymes of cytochrome P450 (CYP450 1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2 and 4A1), fatty acyl-CoA oxidase (FACO) and select Phase II conjugating enzymes (UDPGT, GST and ST) in the rat using specific 5′ and 3′ oligonucleotide primers and reverse transcriptase-polymerase chain reaction. The method closely reproduces the expression obtained from rat liver tissue following treatment with the same enzyme inducers. Constitutive and inducible expression was maintained by resuspending, culturing and then overlaying adult rat hepatocytes with an extracellular matrix such as Matrigel®.

摘要翻译： 肝细胞培养系统，引物组和选择性检测和定量评估细胞色素P450主要同工酶（CYP450 1A1，1A2,2B1 / 2,2C11,2E1,3A2,3A2和4A1）的mRNA表达水平的分析方法，脂肪 酰基辅酶A氧化酶（FACO），并使用特异的5'和3'寡核苷酸引物和逆转录酶 - 聚合酶链反应在大鼠中选择II期缀合酶（UDPGT，GST和ST）。 该方法在用相同的酶诱导剂处理后，紧密地再现了从大鼠肝组织获得的表达。 通过用细胞外基质如Matrigel重悬，培养，然后覆盖成年大鼠肝细胞来保持组成型和诱导型表达。

公开(公告)号：US6127117A

公开(公告)日：2000-10-03

申请号：US855583

申请日：1997-05-13

申请人： Dale Lynn Morris ,  Julio Cesar Davila

发明人： Dale Lynn Morris ,  Julio Cesar Davila

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P19/34

CPC分类号： C12Q1/686 ,  C12Q1/6876 ,  C12Q1/6888 ,  C12Q2600/142 ,  C12Q2600/158

摘要： Hepatocyte culturing system, primer sets and an analytical method for selectively detecting and quantitatively assessing the levels of mRNA expression of the major isoenzymes of cytochrome P450 (CYP450 1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2 and 4A1), fatty acyl-CoA oxidase (FACO) and select Phase II conjugating enzymes (UDPGT, GST and ST) in the rat using specific 5' and 3' oligonucleotide primers and reverse transcriptase-polymerase chain reaction. The method closely reproduces the expression obtained from rat liver tissue following treatment with the same enzyme inducers. Constitutive and inducible expression was maintained by resuspending, culturing and then overlaying adult rat hepatocytes with an extracellular matrix such as Matrigel.RTM..

摘要翻译： 肝细胞培养系统，引物组和选择性检测和定量评估细胞色素P450主要同工酶（CYP450 1A1，1A2,2B1 / 2,2C11,2E1,3A2,3A2和4A1）的mRNA表达水平的分析方法，脂肪 酰基辅酶A氧化酶（FACO），并使用特异的5'和3'寡核苷酸引物和逆转录酶 - 聚合酶链反应在大鼠中选择II期缀合酶（UDPGT，GST和ST）。 该方法在用相同的酶诱导剂处理后，紧密地再现了从大鼠肝组织获得的表达。 通过用细胞外基质如Matrigel TM重悬，培养，然后覆盖成年大鼠肝细胞来维持组成型和诱导型表达。