Mass isolation of pole cells from Drosophila melanogaster☆
[摘要] Crudecellsuspensionswereobtainedfrom3×106pregastrulastagedembryos.ThesecellsfractionatedintothreebandsonRenografindensitygradients.Usingultrastructuralcharacteristicsforidentification,polecellswerelocalizedinthetwodenserbands.EManalysesalsorevealedastrikingdifferenceinthefrequencyoflipidvacuolesfoundinthepole-cellcytoplasmversusthecytoplasmofotherembryoniccells.Thisdifferencehasenabledustoidentifypolecellsbylightmicroscopyusinganeutrallipidstain.ThroughdetailedanalysesoftheRenografinfractionswiththisstain,wehaveshownthatpolecellsresolveintotwotothreedensityclasses.Evidenceispresentedwhichsuggeststhatthesedensityclassesreflectdifferentdevelopmentalageclasses.Thesizedifferencesbetweenthepolecellsandotherembryoniccellscontainedintheenrichedpole-cellfractionsfromthedensitygradientshasenabledustousesedimentationvelocitycentrifugationforadditionalenrichment.Adistinctlowerbandofcellswasobtainedwhichconsistedof80%polecells.Usingtheseprocedures,1–2×107polecellscanbeobtaineddaily.
[发布日期] [发布机构]
[效力级别] [学科分类] 生物科学（综合）
[关键词] [时效性]
