An efficient protocol for deriving liver stem cells from neonatal mice: validating its differentiation potential.

The success of liver regeneration depends on the availability of suitable cell types and their potential to differentiate into functional hepatocytes. To identify the stem cells which have the ability to differentiate into hepatocytes, we used neonatal liver as source. However, the current protocol for isolating stem cells from liver involves enzymes like collagenase, hyaluronidase exposed for longer duration which limits the success. This results in the keen interest to develop an easy single step enzyme digestion protocol for isolating stem cells from liver for tissue engineering approaches. Thus, the unlimited availability of cell type favors setting up the functional recovery of the damaged liver, ensuring ahead success towards treating liver diseases. We attempted to isolate liver stem derived cells (LDSCs) from mouse neonatal liver using single step minimal exposure to enzyme followed by in vitro culturing. The cells isolated were characterized for stem cell markers and subjected to lineage differentiation. Further, LDSCs were induced to hepatocyte differentiation and validated with hepatocyte markers. Finally, we developed a reproducible, efficient protocol for isolation of LDSCs with functional hepatocytes differentiation potential, which further can be used as in vitro model system for assessing drug toxicity assays in various preclinical trials.

Specific targeting of Ep-CAM in various carcinomas by novel monoclonal antibodies.

Epithelial cell adhesion molecule (Ep-CAM) is a 40 kDa transmembrane glycoprotein overexpressed in majority of tumor epithelial cells and has a major morphoregulatory function, relevant not only to epithelial tissue development, but also in carcinogenesis and tumor progression. Since Ep-CAM localizes at the cell surface of most carcinomas, the molecule is an attractive target for immunotherapy and several strategies have been deployed to treat cancer using Ep-CAM targeting, including MAb therapy. For improving effective targeting of this protein for diagnostics in various clinical samples, we generated and characterized an anti-Ep-CAM MAb (C4) using recombinant Ep-CAM protein, comprising the highly immunogenic domain. The specificity of C4-MAb was characterized in Ep-CAM positive cell lines (PC3 and MCF-7) by flow cytometry and immunofluorescence. The immunohistochemistry analysis in clinical tissue samples showed specific detection of epithelial antigens in breast, colon, stomach, and prostate carcinomas. Thus, this Ep-CAM MAb (C4-MAb) could be used for both diagnostic and therapeutic applications due to its specificity.