Production of Functional Hepatobiliary Organoids from Human Pluripotent Stem Cells

The research on human hepatobiliary development and disorders has been constrained by minimal access to human fetal tissue, and low accuracy of animal models. To overcome this problem, we have established a system for the differentiation of human pluripotent stem cells (hPSCs) into functional hepatobiliary organoids (HBOs). We have previously reported that our 45-d approach closely mimics key stages of hepatobiliary development, starting with the differentiation of hiPSC into endoderm and a small part of mesoderm, and subsequently into hepatoblast-like cells, followed by the parallel generation of hepatocyte-like cells and cholangiocyte-like cells, formation of immature HBO expressing early hepatic and biliary markers, and mature HBO displaying hepatobiliary functionality. In this study, we present an updated version of our previous protocol, which only needs 35 days to achieve maturation in vitro. Furthermore, a hepatobiliary culture medium is developed to functionally maintain the HBOs for more than 1.5 months. The capacity of this approach for producing large amounts of functional HBOs and enabling long-term culture in vitro holds promise for applications on developmental research, disease modeling, as well as screening of therapeutic agents.

Preparation and characterization of monoclonal antibody against translationally controlled tumor protein.

From our previous proteomic research, we found that translationally controlled tumor protein (TCTP) might play at least a partial role in colon adenocarcinoma progression. However, the precise impact of TCTP on colorectal cancer metastasis progression is currently still unknown. Therefore, immunology reagents are urgently needed to proceed with the next mechanism-related research. Moreover, the identification of TCTP expression level in tissue of colorectal cancer patients also requires substantial amounts of immunology reagents. In this report, monoclonal antibodies (MAbs) against to TCTP were made from hyperimmune Balb/c mice, by injecting 50 µg of purified antigen intraperitoneally. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using purified protein. Finally six mouse hybridomas producing MAbs to TCTP were established. The MAbs obtained were fully characterized using Western blot analysis and immunohistochemistry. The results showed that these antibodies could be used for the preliminary application of the next mechanism-related research and TCTP expression level analysis.