3D culture of functional human iPSC-derived hepatocytes using a core-shell microfiber
IPSC human hepatocytes derived great promise as a source of cells for cell therapy and drug screening. However, the hepatocyte culture method for highly-quantitative unassigned. Here, we have developed a 3D encapsulation and cultivation methods for IPSC-hepatocytes in microfiber hydrogel core-shell (aka cell fiber).
3D microenvironment in the form of fibers composed of abundant extracellular matrix (ECM), the IPSC-liver hepatocytes exhibited many characteristics, including albumin secretion and gene expression markers of liver (ALB, HNF4α, ASGPR1, CYP2C19, and CYP3A4). In addition, we found that the fibers are mechanically stable and can apply for hepatocyte transplantation.
Three days after transplantation of microfiber into the abdominal cavity immunodeficient mice, human albumin was detected in the peripheral blood of mice transplanted. These results indicate that fiber-hepatocytes promising IPSC well as in vitro models for drug screening or as graft implantation for treating heart failure.
Visualization and isolation of murine hepatocytes special zones which maintain the expression of different cytochrome P450 oxidase in primary culture
parenchymal hepatocytes are responsible for most of the metabolic function of the liver, but showing a different functional properties depending on their localization in the liver lobule. P450 oxidase is a family of drug-metabolism enzyme, which is expressed primarily in hepatocytes local in centrilobular regions (zone 3). This paper describes a unique strain of transgenic mice that distinguishes zone 3 hepatocytes of periportal zone 1 hepatocytes by EGFP fluorescence intensity.
The second zone 1 and zone 3 hepatocytes isolated from these mice showed the zone-specific gene expression patterns were the same as in the liver tissue in vivo.
Experiments using primary cultures of hepatocytes demonstrated that the combination of low oxygen concentration and activation of Wnt / β-catenin expression zones maintain drug-metabolism enzyme 3-specific P450, marked by their susceptibility to acetaminophen-induced mitochondrial dysfunction. Hepatocytes special zone provides a useful system in the pathophysiology of liver research and drug development.
Erythromycin evaluation as a tool to assess the contribution of CYP3A low clearance compound in hepatocyte cultures long term
Long-term cultures of hepatocytes as HepatoPac® system is suitable for evaluating the metabolic turnover low clearance (CL) for sustained drug metabolism capacity and their long-term survival. Erythromycin (ERY), moderate CYP3A inhibitors based mechanisms, evaluated as a tool in HepatoPac models to assess the contribution of CYP3A for clearance drug candidates.
ERY inhibit CYP3A activity by 58 and 80% for the 3 and 10 pM, respectively, up to 72 hours. At 30 pM, ERY MDZ hydroxylation is inhibited by> 85% for the entire duration of the incubation of 144 hours. Clint APZ inhibited 58% by 3 pM ERY, 75% by 15 pM ERY, 89% by 30 pM ERY and 94% by 60 pM ERY. ERY (30 M) did not significantly affect Clint substrate for several other major CYP isoforms was evaluated and no real inhibit UGT isoforms 1A1, 1A3, 1A4, 1A6, 1A9, 2B7 or 2B15 were assessed using recombinant UGTs.
ERY is only slightly increased CYP3A4 gene expression 2.1-fold (14% of RIF induction) at 120 pM indicates that the effective concentration for the inhibition of CYP3A activity (30-60 uM), AHR (arylhydrocarbon receptor), CAR (constitutive androstane receptor ) and PXR (pregnane X receptor) activation is not possible to increase the apparent levels of an enzyme or other metabolic drug transporter.Connection error.
ERY at concentrations up to 60 pM non-toxic to 6 days of incubation. Use of ERY to selectively inhibit CYP3A in hepatocytes, long-term high-functioning as HepatoPac can be a valuable strategy to evaluate the contribution