Abstract
Purpose: :
To verify the isolation method of primary mouse retinal ganglion cells (RGCs) by immunopanning-magnetic separation method.
Methods: :
Retinal cell suspension was obtained from one to four day-old ICR mouse eyeballs. Primary RGCs were isolated using three methods: 1) direct magnetic separation method, 2) two step immunopanning method, and 3) immunopanning-magnetic separation method.
Results: :
As determined by Immunocytochemical staining and Western immunoblots, the primary RGCs isolated using direct magnetic separation method were contaminated by syntaxin-positive retinal amacrine cells; the cells isolated using two step immunopanning method were mixed by GFAP-positive retinal glial cells; the RGCs isolated using immunopanning-magnetic separation method showed the highest purity compared to other methods.
Conclusions: :
Immunopanning-magnetic separation method is a useful method for isolation of primary mouse RGCs in vitro.
Keywords: ganglion cells • retinal culture