4.1 Article

Construct Double Reporter Vector Enhanced by CK19 Promoter and Its Application in Differentiation of Hepatic Progenitor Cells

Journal

PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
Volume 37, Issue 7, Pages 728-736

Publisher

CHINESE ACAD SCIENCES, INST BIOPHYSICS
DOI: 10.3724/SP.J.1206.2010.00034

Keywords

cytokeratin; renilla luciferase; red fluorescence protein; hepatic oval cells; differentiation

Funding

  1. Hi-Tech Research and Development Program of China [2006AA02A107]
  2. National Basic Research Program of China [2005CB522702]
  3. National Natural Science Foundation of China [30901441]

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Liver progenitor cells are co-precursor cells of hepatic cells and bile duct epithelial cells, a reporter gene was used to research the differentiation of liver progenitor cells. First of all, the cytokeratin 19 promoter segment was cloned from hepatocellular carcinoma cell line HepG2 and then following the promoter a renilla luciferase and a red fluorescence protein's fusion gene were inserted to finish the double report lentiviral vector. Second, the liver progenitor cells were transducted with lentivirus, and then GFP positive cells were enriched by flow cytometry sorting. Third, the GFP positive liver progenitor cells were co-cultured with PT67 cells which could express the molecule-epimorphin for 5 days. Then, it was found that the stable transducted liver progenitor cells' shape were not only transformed and arranged as cord like structure, but also renilla luciferase and red fluorescence protein which enhanced by CK19 promoter were detected. So, these results proved that the liver progenitor cells had been induced to bile duct epithelial cells. The vector enhanced by CK19 promoter can monitor the differentiation of liver progenitor cells in different environment. In a word, this lentivirus vector can help us study the differentiation mechanism of liver progenitor cells, and scan the molecules which can do a help in differentiation.

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