Possibility of purifying the Spermatogenic Cells in rat using Gravitational Split-Flow Thin Cell (GrSPLITT) Fractionation

Abstract

The Gravitational split-flow thin cell (GrSPLITT) fractionation is a rapid, gentle and continuous separation technique that should be used to separate spermatogenic cells into three stages: (1) spermatogonia and spermatocyte (2) round spermatids and (3) elongate spermatids and spermatozoa. The different stages of the spermatogenic cells display differences in density and size. In this work, two modes of GrSPLITT namely, full-feed depletion mode (FFD-GrSPLITT) (and transport mode (TS-GrSPLITT) were compared for their possibility to separate the three stages of spermatogenic cells. Imaging and sizing of the original sample and all fractions were analyzed to obtain their size distribution using dark field microscopy with analysis software. The percentage of purity and recovery were calculated and used to objectively compare their separation efficiencies of the two mods of the FFD mode and the TS mode of GrSPLITT fractionations. All experiments showed that the TS-GrSPLITT fractionation trends to be the better separation efficiency than the FFDGrSPLITT fractionation.