Cellular and Molecular Physiology Core Facility

This Core facility provides both traditional and cutting-edge experimental models as resources to Center members. It is organized to provide these resources, along with relevant technical expertise, equipment and personnel to Liver Center investigators who wish to work with animal models of liver disease, isolated liver cells, or gene expression in liver tissue.

The Cellular and Molecular Physiology Core is divided into two components: the cell isolation and cell culture component and the molecular component.

This includes:

• Isolated cell preparations
• Cell culture facilities
• Protein and gene expression
• Altering gene expressing in cells and tissues
• iPSC/liver organoid service

By centralizing procedures in a Core facility, investigators are assured of a high degree of quality control and preparations that can often be used simultaneously by more than one investigator.

This facility isolates hepatocytes and non-parenchymal liver cells (cholangiocytes, endothelial cells, stellate cells, portal fibroblasts and hepatic lymphocytes) primarily from rat and mouse. Human hepatocytes are available periodically from Univ. of Pittsburgh; Interested individuals should contact Mateus Guerra or Emma Kruglov. In addition, the Core maintains frozen stocks of some commonly used cell lines, such as HepG2, which can be provided to Liver Center members.

Expertise is available also for rat and mouse animal models of cholestatic liver disease (bile duct ligation, estrogen administration, LPS administration), fibrosis and cirrhosis (bile duct ligation, CCL4), and liver regeneration (partial hepatectomy). In addition, bile can be collected to measure bile flow in anesthetized mice. Two Matrix Anesthesia Systems are part of the Isolation component, with one available for other investigators within the facility. These have increased the number of animals that can be utilized both for in-vivo studies of bile secretion as well as preparation of isolated liver cell populations.

Available antibodies

The primary role of this component is to provide state-of-the-art molecular equipment and technical expertise and training to Liver Center members in a centralized facility. The equipment below can be reserved by Liver Center members via https://yalelivercenter.zohobookings.com/#/yalelivercenter. New users must be trained by Mateus Guerra.

This component includes:

1. Two real time RT-PCR systems for gene expression, quantification, and genotyping

• QuantStudio 6 Flex qPCR
• QuantStudio 3 qPCR

2. Fotodyne Imaging Workstation: This instrument uses a CCD camera to photograph gels and blots in either transmitted white and UV light or reflected white light modes. Interested individuals should contact Mateus Guerra.

3. BioTek Synergy H1MF: This equipment provides fluorescence intensity, time resolved fluorescence, fluorescence polarization, glow and flash luminescence, UV-Vis absorbance, FRET, TR-FRET, BRET, area scanning, and spectral scanning

4. Beckman L8-70M Ultra centrifuge

5. Access to the Keck Biotechnology Resource Laboratory: The Keck Biotechnology Resource Laboratory provides state-of-the-art biotechnology resources throughout the University. Molecular Sequencing and primer construction are common facilities available to all Yale faculty. The Keck facilities also include advanced biostatistical resources, microarray facilities, and a dedicated proteomics core that are cost-shared with this Center.