Improved protocols for protein and RNA isolation from three-dimensional collagen sandwich cultures of primary hepatocytes.
The sandwich culture is the most widely used long-term culture system for functional primary hepatocytes. Despite its advantages, the currently available protocols for protein and RNA extraction are either time-consuming or contain steps that may skewer the results. This paper describes improved protocols for RNA and protein extraction from sandwich cultures that are easy to perform, require short working time, and use no additional enzymatic reactions that could change the expression profile of the cells. The quality of the RNA is excellent, allowing also applications requiring high purity such as microarrays. In general, the protocols are suited for any cells in 3D collagen culture.
Heidebrecht
F
F
Cell Techniques and Applied Stem Cell Biology, Biocity, University of Leipzig, Germany. felicia.heidebrecht@yahoo.de
Schulz
I
I
Keller
M
M
Behrens
S-E
SE
Bader
A
A
D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen; D022781:Hepatocytes / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification*
D011506:Proteins; D012313:RNA; D003094:Collagen