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  • Development of Hepatocyte-like Cell Derived from Human Induced Pluripotent Stem cell as a Host for Clinically Isolated Hepatitis C Virus.

Development of Hepatocyte-like Cell Derived from Human Induced Pluripotent Stem cell as a Host for Clinically Isolated Hepatitis C Virus.

Current protocols in stem cell biology (2017-08-15)
Khanit Sa-Ngiamsuntorn, Suradej Hongeng, Adisak Wongkajornsilp
ABSTRACT

This unit describes protocols to develop hepatocyte-like cells (HLCs) starting from mesenchymal stem cells (MSCs) as a natural host for hepatitis C virus (HCV). These include the preparation of MSCs from bone marrow, the reprogramming of MSCs into induced pluripotent stem cells (iPSCs), and the differentiation of iPSCs into HLCs. This unit also incorporates the characterization of the resulting cells at each stage. Another section entails the preparations of HCV. The sources of HCV are either the clinically isolated HCV (HCVser) and the conventional JFH-1 genotype. The last section is the infection protocol coupled with the measurement of viral titer. © 2017 by John Wiley & Sons, Inc.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly(vinyl alcohol), 87-90% hydrolyzed, average mol wt 30,000-70,000
Sigma-Aldrich
Periodic Acid-Schiff (PAS) Staining System
Sigma-Aldrich
Rifampicin, ≥95% (HPLC), powder or crystals
Roche
KAPA2G Fast HotStart ReadyMix, 2 ×
Sigma-Aldrich
Alkaline Phosphatase Detection Kit, This Alkaline Phosphatase Detection Kit is a specific & sensitive tool for the phenotypic assessment of Embryonic Stem (ES) cell differentiation by the determination of AP activity.