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Isolation of CD133+ Liver Stem Cells for Clonal Expansion
Published on: October 10, 2011
Protocolo para el aislamiento optimizado de células mononucleares de tejido hepático y tumoral mediante digestión
Sachin Kumar Singh Chauhan1, Linda Feldbrügge2, Moritz Schmelzle2
1Department of Gastroenterology, Hepatology, Infectious Diseases and Endocrinology, Hannover Medical School, 30625 Hannover, Lower Saxony, Germany.
Abstract:
Mononuclear cells from human liver tissue, including tumor and non-tumor regions, are a valuable source for studies of the tumor microenvironment. Here, we present a protocol to dissociate liver tissue from patients with hepatocellular carcinoma. We provide steps to generate single-cell suspension using either mechanical or enzymatic digestion, following steps for separating mononuclear cells using density gradient. Finally, we describe steps for flow cytometry staining of isolated immune cells. For complete details on the use and execution of this protocol, please refer to Moreno-Fernandez et al.1 and Heinrich et al.2.

