Separation of mouse spermatogenic cells by sedimentation velocity: A morphological characterization☆

[摘要] Amethodutilizingsequentialenzymaticincubationincollagenase(1mg/ml)andtrypsin(2.5mg/ml)hasbeendevelopedforthedissociationoftheseminiferousepithelium.Asignificantadvantageofthismethodisthat,followingcollagenaseincubationandwashingsinanenrichedKrebs-Ringerbicarbonatebuffersolution,isolatedseminiferoustubulesareobtainedwhicharefreeofinterstitialcells.The“purified”seminiferousepitheliumisthendissociatedwithtrypsin.Afurtheradvantageofthisdissociationtechniquehasbeenareductioninthenumberofsymplasts(multinucleatecells)whichformbytheopeningupoftheintercellularbridgesthatoccurbetweensynchronouslydifferentiatingclustersofgermcells.Boththeeliminationoftheinterstitialcellsandthereductioninthenumberofsymplastshavemadepossibletherecoveryofmorehighlyenrichedgermcellfractions.Thehomogeneityofthecellfractionswasdeterminedbylightandelectronmicroscopy.IntegrityoftheisolatedcellswasverifiedbyTrypanblueexclusionandmeasurementofoxygenconsumption.

[学科分类] 生物科学（综合）