TY - JOUR
T1 - Isolation, culture and genetic manipulation of mouse pancreatic ductal cells
AU - Reichert, Maximilian
AU - Takano, Shigetsugu
AU - Heeg, Steffen
AU - Bakir, Basil
AU - Botta, Gregory P.
AU - Rustgi, Anil K.
PY - 2013/7
Y1 - 2013/7
N2 - The most common subtype of pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). PDAC resembles duct cells morphologically and, to some extent, at a molecular level. Recently, genetic-lineage labeling has become popular in the field of tumor biology in order to study cell-fate decisions or to trace cancer cells in the mouse. However, certain biological questions require a nongenetic labeling approach to purify a distinct cell population in the pancreas. Here we describe a protocol for isolating mouse pancreatic ductal epithelial cells and ductlike cells directly in vivo using ductal-specific Dolichos biflorus agglutinin (DBA) lectin labeling followed by magnetic bead separation. Isolated cells can be cultured (in two or three dimensions), manipulated by lentiviral transduction to modulate gene expression and directly used for molecular studies. This approach is fast (~4 h), affordable, results in cells with high viability, can be performed on the bench and is applicable to virtually all genetic and nongenetic disease models of the pancreas.
AB - The most common subtype of pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). PDAC resembles duct cells morphologically and, to some extent, at a molecular level. Recently, genetic-lineage labeling has become popular in the field of tumor biology in order to study cell-fate decisions or to trace cancer cells in the mouse. However, certain biological questions require a nongenetic labeling approach to purify a distinct cell population in the pancreas. Here we describe a protocol for isolating mouse pancreatic ductal epithelial cells and ductlike cells directly in vivo using ductal-specific Dolichos biflorus agglutinin (DBA) lectin labeling followed by magnetic bead separation. Isolated cells can be cultured (in two or three dimensions), manipulated by lentiviral transduction to modulate gene expression and directly used for molecular studies. This approach is fast (~4 h), affordable, results in cells with high viability, can be performed on the bench and is applicable to virtually all genetic and nongenetic disease models of the pancreas.
UR - http://www.scopus.com/inward/record.url?scp=84904650619&partnerID=8YFLogxK
U2 - 10.1038/nprot.2013.079
DO - 10.1038/nprot.2013.079
M3 - Article
C2 - 23787893
AN - SCOPUS:84904650619
SN - 1754-2189
VL - 8
SP - 1354
EP - 1365
JO - Nature Protocols
JF - Nature Protocols
IS - 7
ER -