Back to Search
Start Over
Preparation of Kupffer cell enriched non-parenchymal liver cells with high yield and reduced damage of surface markers by a modified method for flow cytometry.
- Source :
-
Cell Biology International . Apr2013, Vol. 37 Issue 4, p284-291. 8p. - Publication Year :
- 2013
-
Abstract
- The aim of this study was to optimise a collagenase perfusion protocol for the isolation of a liver non-parenchymal cell (NPC) suspension enriched for Kupffer cells that reduced damage to F4/80 antigen cell surface expression to allow analysis by flow cytometry. Kupffer cell-enriched liver NPCs were isolated from C57BL/6 mice using different protocols. Flow cytometry was used to examine the effect of collagenase digestion on F4/80 expression on Kupffer cells, and results were represented by the percentage of F4/80 positive cells and by the F4/80 mean fluorescence intensity (MFI). The perfusion temperature, concentration of collagenase solution and total dosage of collagenase for liver perfusion influenced the effect of collagenase perfusion on the expression of F4/80 antigen on Kupffer cells. Collagenase perfusion at 28°C resulted in an increased percentage of F4/80 positive cells ( P = 0.001) and MFI ( P = 0.005) compared with 37°C. Perfusion with a total dose of 1.0 g/kg BW collagenase (using a 0.75 mg/mL solution) resulted in the highest percentage of F4/80 positive cells ( P = 0.001) compared with 0.8 g/kg BW and 1.2 g/kg BW collagenase. Isolation of cells using the modified protocol resulted in a higher percentage of Kupffer cells ( P < 0.001) and a higher MFI of F4/80 antigen ( P < 0.001) compared with the common protocol. [ABSTRACT FROM AUTHOR]
- Subjects :
- *KUPFFER cells
*COLLAGENASES
*LIVER cells
*FLOW cytometry
*ANTIGEN presenting cells
Subjects
Details
- Language :
- English
- ISSN :
- 10656995
- Volume :
- 37
- Issue :
- 4
- Database :
- Academic Search Index
- Journal :
- Cell Biology International
- Publication Type :
- Academic Journal
- Accession number :
- 101074399
- Full Text :
- https://doi.org/10.1002/cbin.10035