基于多聚鸟氨酸海藻酸微囊化人正常肝细胞系的体外培养-论文.pdf

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1、北京医学2013年第35卷第11期·951·．论著．基于多聚鸟氨酸海藻酸微囊化人正常肝细胞系的体外培养王健冯晓明柯林楠王春仁【摘要l目的观察海藻酸钠一多聚鸟氨酸一海藻酸(alginate—PLO—alginate，A—PLO—A)微囊和海藻酸钡一多聚鸟氨酸一海藻酸(Ba—alginate—PLO—alginate，B—PLO—A)两种微囊包裹人正常肝细胞系HL一7702细胞后．肝细胞形态和功能的变化，评价PLO对微囊化肝细胞的生物学行为的影响。方法采用优化的微囊制备条件，制备A—PLO—A微囊、B—PLO—A微囊

2、．并包裹HL一7702细胞，观察肝细胞在基于多聚鸟氨酸的微囊中的生长增殖的情况及代谢产物浓度水平。结果A—PLO—A和B—PLO—A两种微囊包裹肝细胞后，体外培养1周．细胞均可在两种微囊内形成不同大小和形态的肝细胞聚集体，微囊光滑无破损现象，台盼兰染色计数的结果显示，包囊前细胞存活率为94％；A—PLO—A微囊包囊后细胞存活率为92％，B—PLO—A微囊包囊后细胞存活率为9l％。两种微囊化HL一7702细胞各时间点ALB、BUN分泌量差异均无统计学意义结论微囊囊膜的存在并未对具有贴壁生长和接触抑制性特征的HL一7

3、702细胞的增殖和生理功能产生明显的不利影响基于多聚鸟氨酸的新型海藻酸微囊在一定程度上可以满足微囊化肝细胞生存的需要．并可维持其正常的生理功能【关键词】多聚鸟氨酸微囊肝细胞组织工程Humannormallivercelllinecultureinvitrobasedonpoly-ornithineonalginatemicroeneapsulatedteniqueWANGJian，FENGXiao—ruing，KELin—nan，etal(NationalInstituteforFoodandDrugContro

4、l，Beijing100050)【Abstract】ObjectiveToobservethemorphologyandphysiologicalfunctionofnormallivercellsculturedinvitrobasedonpoly—ornithineonalginatemicroencapsulatedtenique．MethodsInthisstudy，themicrocapsulepreparationconditionsonalginate-polyornithine—alginate(

5、A-PLO-A)microcapsulesandbariumalginate-polyoruithine—alginate(B—PLO—A)microcapsuleswereoptimizedfirstandmicroencapsulated．Thenthemorphology，metaboliteconcentra—tionlevelsofHL一7702cellsinvitroweretestedandthegrowthaswellproliferationoflivercellsinpoly-or-nithi

6、nemierucapsuleweremonitored．ResultsItshowedthatafter1weekofinvitroculture，thetwomicr0encapsulatedhepatocytes，A-PLO-AandB-PLO—A，couldgrowandformlivercellaggregatesinvarioussizesandmorphology．Noevi—dentmorphologychanges，ceilssurroundingandfibrosiswereobservedin

7、thecollectedmicrocapsules．AndthetwomicroencapsulatedHL一7702cellsbothhadthecapabilityofpersistentsynthesisofproteinandurea．Therewasnosig—nificantdifferenceintheconcentrationlevelbetweenthetwosetsofcells．ConclusionThepresenceofmicrocapsulemembranedonotnegativee

8、fectontheproliferationandbiologicalfunctionofHL-7702ceilswhichcarrythenatureofadherentandcontactinhabitationcharacteristics．Thishasprovedthatpoly—omithineonthenewalgi—natemicrocapsulesmay