Rat retinal ganglion cells culture enriched with the magnetic cell sorter

The magnetic cell sorter (MACS) technique was applied to isolate retinal ga nglion cells (RGCs) for culture. RGCs were labeled retrogradely with 1.1'-d ioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). Subsequ ently retinal cell suspensions were incubated with biotinylated anti-rat Th y-1 antibody and MACS Streptavidin MicroBeads, and then applied onto the co lumn in the magnetic fields. Cells attached on the column were flashed out without magnetism and plated on glass cover slips. RGCs were enriched to 31 .0% of all cells with MACS from 0.55% before applying onto the magnetic col umn. Mean diameters of DiI-labeled cells were significantly larger than tho se of unlabeled cells. All cells with soma diameter over 11 mu m were label ed. The number of viable RGCs were counted in the 10 fields of six cultures at a magnification of x200; the mean numbers on the 2nd, 7th and 14th cult ure-day were 53 +/- 3, 24 +/- 2 and 21 +/- 3, respectively (mean +/- SEM, n = 6). Thus, the MACS technique was confirmed to be useful for enrichment o f RGCs and long-term study of cultured RGCs. (C) 1999 Elsevier Science irel and Ltd. All rights reserved.