Clinically relevant CYP3A4 induction mechanisms and drug screening in 3D spheroid cultures of primary human hepatocytes.
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Clinically relevant CYP3A4 induction mechanisms and drug screening in 3D spheroid cultures of primary human hepatocytes.
CYP3A4 induction is an important cause of drug-drug interactions, making early identification of drug candidates with the obligations CYP3A4 induction in drug development prerequisites. Here, we present a 3D spheroid cultures of primary human hepatocytes (PHH) as CYP3A4 induction of new screening models.
Screening of 25 drugs (12 known inducers of CYP3A4 in vivo and 13 negative control) at physiologically relevant concentrations revealed 100% sensitivity and 100% specificity system. Three of inducers of CYP3A4 in vivo induction of CYP3A4 displayed capacity is much higher in comparison with the 3D spherical culture in 2D monolayer culture. Among them, we identify AZD1208, PIM kinase inhibitor in Phase I development stopped because of unexpected autoinduction CYP3A4, CYP3A4 inducer as the only active in 3D spheroids but not in 2D monolayer culture.
Gene knockdown experiments revealed that AZD1208 require PXR to induce CYP3A4. Rifampicin require PXR solely to induce CYP3A4 and CYP2B6, while phenobarbital-mediated induction of CYPs is not indicative of the absolute dependence on both PXR or CAR demonstrated its ability to switch the activation of nuclear receptors. Mechanistic Study of AZD1208 to find involvement of MAPK / ERK pathway in induction of CYP3A4-sensitive culture format used, as revealed by the inhibition of ERK1 / 2 phosphorylation Tyr204 and sensitivity to EGF pressure. Accordingly, we also identified lapatinib, a dual EGFR / HER2 inhibitor, such as CYP3A4 inducer active just another round in 3D culture.
Our findings offer insight into the pathways involved in the induction of CYP3A4 and suggest spheroids PHH for preclinical screening of CYP3A4 induction.
Clinically relevant CYP3A4 induction mechanisms and drug screening in 3D spheroid cultures of primary human hepatocytes.
Glucose and lipid metabolism screening models of hepatocyte spheroids after culture by injecting fiber fragments.
With the increase in obesity, diabetes and other metabolic diseases, in vitro liver cell and tissue models play an important role in the identification of active pharmaceutical ingredients. Up to now three-dimensional (3D) models rarely focuses on the culture of hepatic glucose and lipid metabolism.
In addition, primary human liver cells suffer from limited availability and differences among donors to build the model reproduced. Thus, in this study, the most available human liver cancer cells (HepG2), and primary hepatocytes from rat (RPH) proposed to build a 3D spheroids using fiber fragments graft syringe with galactose (GSF) as a substrate. RPH and spheroids showed HepG2 cells and cell fragments strong-fiber interaction to promote cell survival, albumin and urea synthesis.
Compared with HepG2 spheroids, spheroids RPH demonstrate strong abilities in terms of consumption of glucose metabolism of glucose, glycogen intracellular content, the level of glucose in gluconeogenesis and sensitivity to the hormone modulators such as insulin and glucagon. On the other hand, HepG2 spheroids displays a strong ability of lipid metabolism in produce significantly higher levels of total cholesterol and triglycerides.
Compared with those who did not fragment the fiber, the GSF-supported 3D culture establish an effective model for in vitro glucose (RPH spheroids) and lipid metabolism (HepG2 spheroids). The model confirmed screening of each enzyme activity and gene expression and showed significantly higher sensitivity and clinically relevant response to hypoglycemic and lipid-lowering drugs.
Hepatocytes of male Wistar rat in monolayer, in 12 well-plate
Description: Gentaur offers an assortment of various human and non-human hepatic derived cells. These include Hepatocytes, Total Liver Cell Population (TLC), Stellates, Progenitors and Intra-hepatic biliary epithelial cells. We offer these as cryopreserved cells for convenience. Cryopreserved cells are suitable for a variety of assays including induction, toxicity, drug metabolism and systems biology. Both adherent and suspension cells are available. Custom configurations are available upon request.
Liver Dissociation System 2 (Hepatocytes, Parenchymal hepatocytes), Mouse and Rat
Description: The liver fulfills many vital processes in mammals. It is the central organ of energy metabolism, biotransformation of xenobiotics, and synthesis of plasma proteins under physiological and pathophysiological conditions. Primary human hepatocyte culture is an important experimental model for the study of liver specific function. Propagation of human hepatocytes is critical for gene therapy, cell transplantation and clinical liver support systems. In appropriate culture condition, cultured human hepatocytes proliferate and maintain differentiated hepatocyte function such as the synthesis of serum proteins. HH from Gentaur Research Laboratories are isolated from human liver. HH are cryopreserved immediately after purification and delivered frozen. Each vial contains 1 x 10^6 cells in 1 ml volume. HH are characterized by immunofluorescent method with antibodies to albumin, cytokeratin-18 and vimentin. HH are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast and fungi. HH is not recommended for expanding or long term cultures since the cells do not proliferate in culture.
Description: Our High Quality Mouse Hepatocytes have been isolated from mouse livers after liver perfusion and low-speed differential centrifugation. The hepatocytes are cryoperserved immediately after the experimental process and delivered frozen.
Description: Gentaur offers an assortment of various human and non-human hepatic derived cells. These include Hepatocytes, Total Liver Cell Population (TLC), Stellates, Progenitors and Intra-hepatic biliary epithelial cells. We offer these as cryopreserved cells for convenience. Cryopreserved cells are suitable for a variety of assays including induction, toxicity, drug metabolism and systems biology. Both adherent and suspension cells are available. Custom configurations are available upon request.
Description: The cell line was prepared by immortalizing hepatocytes from a 12 year-old Caucasian female donor with the SV40 large T antigen. The donor tested negative for CMV, HIV, HBV, and HCV.
Description: Gentaur offers an assortment of various human and non-human hepatic derived cells. These include Hepatocytes, Total Liver Cell Population (TLC), Stellates, Progenitors and Intra-hepatic biliary epithelial cells. We offer these as cryopreserved cells for convenience. Cryopreserved cells are suitable for a variety of assays including induction, toxicity, drug metabolism and systems biology. Both adherent and suspension cells are available. Custom configurations are available upon request.
Description: Freshly isolated human hepatocytes represent a living biological system in which to measure metabolism, induction, toxicity, and transporter studies.
Description: Human Pooled Primary Hepatocytes (Heps) provide an ideal culture model for drug testing and drug development.Heps 20 Human Pooled Primary Hepatocytes contain cryopreserved hepatocytes pooled from 20 different donors and have undergone only one freeze cycle. Pooled hepatocytes provide the genetic diversity necessary for superior reliability and predictability of the metabolic characteristics of a test compound relative to a representative population. Pooled hepatocytes increase experimental control and reduce costs by eliminating the need to conduct redundant experiments necessary to obtain population-based data. Each lot has undergone a single freeze cycle, assuring maximum hepatocyte viability and consistency from donor to donor within the pool.Cell Features:Each donor's hepatocytes have been cryopreserved ONCE immediately after isolation and purification.Pools of 10 or 20 donors are available. Each donor is represented by a statistically significant number of cells relative to the pool.There is extensive quality control providing characterization of viability and enzyme activity from both individual donors and the total pool.Gentaur guarantees performance and quality.Donor information is available.
Description: Human Pooled Primary Hepatocytes (HP2 Heps) provide an ideal culture model for drug testing and drug development. HP2 10 Human Pooled Primary Hepatocytes contain cryopreserved hepatocytes pooled from 10 different donors and have undergone only one freeze cycle. Pooled hepatocytes provide the genetic diversity necessary for superior reliability and predictability of the metabolic characteristics of a test compound relative to a representative population. Pooled hepatocytes increase experimental control and reduce costs by eliminating the need to conduct redundant experiments necessary to obtain population-based data. Each lot has undergone a single freeze cycle, assuring maximum hepatocyte viability and consistency from donor to donor within the pool.Cell Features:Each donor's hepatocytes have been cryopreserved ONCE immediately after isolation and purification.Pools of 10 or 20 donors are available. Each donor is represented by a statistically significant number of cells relative to the pool.There is extensive quality control providing characterization of viability and enzyme activity from both individual donors and the total pool.Gentaur guarantees performance and quality.Donor information is available.
Description: Gentaur's hepatocytes for suspension assays (HP1 Heps-s - single donor, male) provide an economic source of characterized human hepatocytes for use in short-term suspension assays to evaluate the metabolism of drug candidates. Gentaur's hepatocytes are a functional, convenient and economical solution for your drug testing and basic research needs. Each lot is provided with extensive characterization of metabolic functionality to ensure that you receive an ideal model for drug testing, drug development or other metabolic studies. Gentaur's human hepatocytes eliminate the need and expense of obtaining liver tissue, and conducting multiple cell isolations; and Gentaur also provides hepatocyte-tested culture reagents and instructions necessary for thawing and culturing your hepatocytes.Cell Features:Suitable for induction assays or metabolic characterization assays in plated or suspension cultures. Plated cultures are excellent for long-term intrinsic clearance assays.Hepatocytes are cryopreserved ONCE immediately after isolation and purification.Characterization of viability and enzyme activity, including fold-induction of CYP1A2, CYP2B6 and CYP3A4 for HP1 Heps-i.Gentaur guarantees performance and quality.Donor demographics and medical information are available.
Description: Gentaur's hepatocytes for suspension assays (HP1 Heps-s, single donor, female) provide an economic source of characterized human hepatocytes for use in short-term suspension assays to evaluate the metabolism of drug candidates. Gentaur's hepatocytes are a functional, convenient and economical solution for your drug testing and basic research needs. Each lot is provided with extensive characterization of metabolic functionality to ensure that you receive an ideal model for drug testing, drug development or other metabolic studies. Gentaur's human hepatocytes eliminate the need and expense of obtaining liver tissue, and conducting multiple cell isolations; and Gentaur also provides hepatocyte-tested culture reagents and instructions necessary for thawing and culturing your hepatocytes.Cell Features:Suitable for induction assays or metabolic characterization assays in plated or suspension cultures. Plated cultures are excellent for long-term intrinsic clearance assays.Hepatocytes are cryopreserved ONCE immediately after isolation and purification.Characterization of viability and enzyme activity, including fold-induction of CYP1A2, CYP2B6 and CYP3A4 for HP1 Heps-i.Gentaur guarantees performance and quality.Donor demographics and medical information are available.
Description: Mouse Hepatocyte Stem Cells; Frozen Vial and Plated cells are available. Cells are only guaranteed with purchase of Gentaur Media and Gentaur Extra Cellular Matrix for appropriate cell culture, for 30 days from the date of shipment.
Description: Gentaur offers an assortment of various human and non-human hepatic derived cells. These include Hepatocytes, Total Liver Cell Population (TLC), Stellates, Progenitors and Intra-hepatic biliary epithelial cells. We offer these as cryopreserved cells for convenience. Cryopreserved cells are suitable for a variety of assays including induction, toxicity, drug metabolism and systems biology. Both adherent and suspension cells are available. Custom configurations are available upon request.
Description: These hepatocytes overcome the functionality, reproducibility, and availability limitations of existing primary cell and tumor-derived cell models by providing the following:Relevant hepatic functionality and phenotypic stabilityExpression of clinical targets that are relevant for liver and metabolic diseaseCompatibility with a wide range of 3D and microphysiological systemsQuality-controlled manufacturing and batch reproducibilityReady supply in a cryopreserved format
Description: Gentaur offers an assortment of various human and non-human hepatic derived cells. These include Hepatocytes, Total Liver Cell Population (TLC), Stellates, Progenitors and Intra-hepatic biliary epithelial cells. We offer these as cryopreserved cells for convenience. Cryopreserved cells are suitable for a variety of assays including induction, toxicity, drug metabolism and systems biology. Both adherent and suspension cells are available. Custom configurations are available upon request.
Description: Gentaur's inducible hepatocytes (HP1 Heps-i, single donor, male) can be plated and used in long-term drug induction studies or metabolic studies where long-term incubations are required. Gentaur's cryopreserved primary human hepatocytes are a functional, convenient and economical solution for your drug testing and basic research needs. Each lot is tested for plating and provided with extensive characterization of metabolic functionality to ensure that you receive an ideal model for drug testing, drug development or other metabolic studies. Gentaur's human hepatocytes eliminate the need and expense of obtaining liver tissue, and conducting multiple cell isolations; and Gentaur also provides hepatocyte-tested culture reagents and instructions necessary for thawing and culturing your hepatocytes.Cell Features:Suitable for induction assays or metabolic characterization assays in plated or suspension cultures. Plated cultures are excellent for long-term intrinsic clearance assays.Hepatocytes are cryopreserved ONCE immediately after isolation and purification.Characterization of viability and enzyme activity, including fold-induction of CYP1A2, CYP2B6 and CYP3A4 for HP1 Heps-i.Gentaur guarantees performance and quality.Donor demographics and medical information are available.
Description: Gentaur's inducible hepatocytes (HP1 Heps-i, single donor, female) can be plated and used in long-term drug induction studies or metabolic studies where long-term incubations are required. Gentaur's cryopreserved primary human hepatocytes are a functional, convenient and economical solution for your drug testing and basic research needs. Each lot is tested for plating and provided with extensive characterization of metabolic functionality to ensure that you receive an ideal model for drug testing, drug development or other metabolic studies. Gentaur's human hepatocytes eliminate the need and expense of obtaining liver tissue, and conducting multiple cell isolations; and Gentaur also provides hepatocyte-tested culture reagents and instructions necessary for thawing and culturing your hepatocytes.Cell Features:Suitable for induction assays or metabolic characterization assays in plated or suspension cultures. Plated cultures are excellent for long-term intrinsic clearance assays.Hepatocytes are cryopreserved ONCE immediately after isolation and purification.Characterization of viability and enzyme activity, including fold-induction of CYP1A2, CYP2B6 and CYP3A4 for HP1 Heps-i.Gentaur guarantees performance and quality.Donor demographics and medical information are available.
Thus, the culture configuration shows the predicted in vitro platform for defining the profiles of glucose and lipid metabolism and screening of therapeutic agents for metabolic disorders such as diabetes and obesity.
