Microencapsulation Technology A Powerful Tool for Integrating Expansion and Cryopreservation of Human Embryonic Stem Cells 英文参考文献.docVIP

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Microencapsulation Technology A Powerful Tool for Integrating Expansion and Cryopreservation of Human Embryonic Stem Cells 英文参考文献.doc

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Microencapsulation Technology A Powerful Tool for Integrating Expansion and Cryopreservation of Human Embryonic Stem Cells 英文参考文献

MicroencapsulationTechnology:APowerfulToolfor IntegratingExpansionandCryopreservationofHuman EmbryonicStemCells MargaridaSerra1,2.,Cla′udiaCorreia1,2.,RitaMalpique1,2,CatarinaBrito1,2,JanneJensen3 ,Petter Bjorquist3,ManuelJ.T.Carrondo2,4,PaulaM.Alves1,2 * 1Instituto de Tecnologia Qu?′mica e Biolo′gica, Universidade Nova de Lisboa, Oeiras, Portugal, 2Instituto de Biologia Experimental e Tecnolo′gica, Oeiras, Portugal, 3CellartisAB,Gotemborg,Sweden,4FaculdadedeCie?nciaseTecnologia,UniversidadeNovadeLisboa,MontedaCaparica,Portugal Abstract The successful implementation of human embryonic stem cells (hESCs)-based technologies requires the production of relevant numbers of well-characterized cells and their efficient long-term storage. In this study, cells were microencapsulated in alginate to develop an integrated bioprocess for expansion and cryopreservation of pluripotent hESCs.Differentthree-dimensional(3D)culturestrategieswereevaluatedandcompared,specifically,microencapsulationof hESCsas:i)singlecells,ii)aggregatesandiii)immobilizedonmicrocarriers.Inordertoestablishascalablebioprocess,hESC- microcapsules were cultured in stirred tank bioreactors. The combination of microencapsulation and microcarrier technologyresultedinahighlyefficientprotocolfortheproductionandstorageofpluripotenthESCs.Thisstrategyensured highexpansionratios(anapproximatelytwenty-foldincreaseincellconcentration)andhighcellrecoveryyields(.70%) aftercryopreservation.Whencomparedwithnon-encapsulatedcells,cellsurvivalpost-thawingdemonstratedathree-fold improvement without compromising hESC characteristics. Microencapsulation also improved the culture of hESC aggregates by protecting cells from hydrodynamic shear stress, controlling aggregate size and maintaining cell pluripotency for two weeks. This work establishes that microencapsulation technology may prove a powerful tool for integratingtheexpansionandcryopreservationofpluripotenthESCs.The3Dculturestrategydevelopedhereinrepresentsa signifi