Production And Cryopreserved Of Microencapsulation Cells And The Culture Of Microencapsulation Embryonic Stem Cells

Microencapsulation technology has been widely used in the study of cell culture andimmunoisolation . APA microcapsules get the more emphasis of researcher for its goodbiocompatibility.While the APAmicrocapsules's making methods have many deficient.Embryonicstem cell is the main solution to the problem the lacking of cell supplication,it is important for celltherapy of future,but the production of embryiod is low during the course of embryonic stem cell's differentiation.. The other problems need be solved instantly are themicroencapsulated cellscryopreservation and transportion.In order to solve the above problems this paper study the Part One Objective: To investingate the probability of producing microcapsules by the selfmade high voltage pulse electrostatic droplet generator. Methods To produce microcapsules byself made high voltage pulse electrostatic droplet generator and determine the size distribution ofmicrocapsules. To compare the effects of different rea -ction time of poly-lysine and sodiumcitromalic acid on microcapsules's diameter and durability. To determine th -eimmunocompatibility and biocompatibility of microcapsules. Results The diameter of themicrocapsules which produced by the device range from 100 micrometer to 750 micrometer.Microcapsule's diameter increases with the increasing of polylysine treating time . Treated withsodium citromalic acid increases the microcapsule size. In vi -tro experiment revealsmicrocapsules has long time durability.It can keep integrity ratio still up to 73.5% in 150thday .The recovery ration is 80% in fifth week after transplant into rat peritoneal cavity .Cells cankeep activity and grow well encapsulated in microcapsules. Conclusion The self-made highvoltage pulse electrostatic droplet gener -ator can produce perfectly sphericall smooth and uniformcapsules which has long durability and good biocompati -bility. Part Two Objective To compare the cryopreservational effects of different type ofmicrocapsules on cells and select the fitful cryoprotectants for microencapsulation.Methods Toencapsulate HK-2 cells by liquefied alginate-polylyssine -alginate microcapsules andalginate-calcium microbeads which produced by self-made high voltage electrostatic dropletgenerator.2 DMSO and glycerol as the cryoprotectants to cryopreserve unencapsulation cells andencapsulation cells.3 to evaluate the defrozed cell's viability by MTT and to determine the proteincontents and DNA contents respectively by coomassie brilliant blue staine and fluorochromeHochest 33258.Results the viability ,protein contents,DNA contents of liquefied microcapsule'scells are significance higher than That of cells encapsulated by microbeads(P<0.05),but there areno significance difference between liquefied microcapsule's cells and unencapsulated cells(P>0.05). The cell viability ,protein contents of the cells which DMSO as cryoprotectants aresignificance higher than that of Glycrerol(P<0.05) but there are no significance differencebetween them on DNA contents(P>0.05). The liquefied microcapsule coupled with DMSO tocryopreserve cells is better than the liquefied microcapsule coupled with glycerol on the cellviability ,protein contents,DNA contents. The liquefied microcapsules are more stable thanmicrobead after be defrozed. Conclusions The cryopreserved effect of the liquefied microcapsulesis better than microbeads and DMSO is more fitful for microcapsules than Glycerol. Part Three objective :to determine the probability of differentiation of embryonic stemcells(ES) into embryonic embryoid(EB) in alginate microcapsules. Methods:to encapsulate theembryonic stem cells marked by green fluorescent protein into alginate microcapsules made byself-made high voltage pulse electrostatic droplet generator and see whether ES can differentiateinto EB .To release the embryoid from microcapsules and calculate the efficiency of ESdifferentiated into EB. To culture the released EB in attachment...