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Developing cryopreservation for Picea sitchensis (sitka spruce) somatic embryos: a comparison of vitrification protocols.
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2008
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FertilityBotanyGeneticsReproductive BiologyEmbryologyReproductive BiotechnologyEmbryo CultureRegenerative MedicinePublic HealthRecovery ResponsesSomatic EmbryosBlastemaMorphogenesisEmbryonic DevelopmentVitrification-based Cryopreservation ProtocolsHuman ReproductionBiologyDevelopmental BiologyBiotechnologyPicea SitchensisVitrification ProtocolsMedicinePlant PhysiologyEmbryonic Stem Cell
Two vitrification-based cryopreservation protocols, encapsulation/dehydration and PVS2 were applied to Stage 2 (globular) and Stage 4 (torpedo) somatic embryos (SE) from Picea sitchensis. Two recovery responses: partially differentiated embryogenic suspensor masses (ESM) and dedifferentiated non-embryogenic masses (NEM) were observed following exposure to LN. All genotypes tested, proliferated NEM, approximately 10 to 100% of the total SE cryopreserved. A General Linear Model applied to NEM recovery data demonstrated several different factors (developmental state and genotype, treatment, culture age) interacted at a significant level (P less than 0.05) to influence proliferation. One genotype was capable of proliferating ESM after cryopreservation using encapsulation-dehydration, this response was achieved for Stage 4 embryos derived from the youngest ESM tissue.