By Takahiro Ochiya
Liver Stem Cells sheds gentle on stem cellphone know-how, that may bring about the improvement of potent medical modalities for human liver ailments. particular subject matters comprise hepatic progenitor cells, hepatic differentiation shape stem cells, bile duct phone formation from stem cells, liver stem cells and extra.
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A small amount of 1 N NaOH was added if necessary. After complete dissolution, the solution was cooled down. If necessary, it was filtrated. 3. 4) added to adjust the total volume to 1 L. This solution can be stored at 4°C for at least 1 year. 3. Isolation of EpCAM+ Cells from Livers of Normal and DDC Diet-Fed Mice 1. Perfusion solution I: Liver Perfusion Medium (GIBCO 17701038). 2. 22-μm STERICUP (Millipore)]. 3. 22-μm STERICUP (Millipore) (made fresh as required) (see Note 1). 4. 22-μm STERICUP (Millipore) (made fresh as required).
Furthermore, we developed culture system and characterized EpCAM+ cells isolated from normal as well as injured livers. Here, we describe the methods for identifying and isolating mouse liver stem/progenitor cells. 2. 1. Generation of Oval Cells in Mouse Liver 1. Adult male mice (8–12 weeks old). C57BL/6 mice were used for all experiments. 2. 1% DDC (Bio-Serv). 2. Immunohistochemistry of Frozen Liver Sections by Zamboni’s Fixation (A Modified ParaformaldehydeBased Fixation Method) Identification and Isolation of Adult Liver Stem/Progenitor Cells 27 1.
6 mM NaHCO3, 14 mM glucose, 100 nM dexamethasone, 28 M. Tanaka and A. Miyajima 1× ITS (insulin, transferrin, selenium X) (GIBCO) and 50 μg/mL gentamicin. 2. Cytokines: Human EGF and human recombinant HGF (final concentration of 10 ng/mL each). 3. Type-I collagen-coated dish: 35 mm dishes were coated with Type-I collagen solution (Cellmatrix Type I-C, Nitta gelatin). 4. 25% Trypsin–EDTA (#25200)(GIBCO). 3. 1. Generation of Oval Cells and Immunohistochemistry 1. Mice were fed by the DDC-containing diet for more than 4 weeks (see Note 2).