微生物学免疫学进展
微生物學免疫學進展
미생물학면역학진전
PROGRESS IN MICROBIOLOGY AND IMMUNOLOGY
2013年
6期
25-29
,共5页
李红芳%赵祖波%任红卫%杨淼%马超%辛军%龚健%李守丽%于滢%徐枫
李紅芳%趙祖波%任紅衛%楊淼%馬超%辛軍%龔健%李守麗%于瀅%徐楓
리홍방%조조파%임홍위%양묘%마초%신군%공건%리수려%우형%서풍
细胞工厂%原代地鼠肾细胞%狂犬病病毒aG株%收获
細胞工廠%原代地鼠腎細胞%狂犬病病毒aG株%收穫
세포공엄%원대지서신세포%광견병병독aG주%수획
Cell factory%Primary Hamster kidney cell%Rabies aG strain%Harvests
目的：为提高生产效率、增加原代地鼠肾细胞单产量及狂犬病病毒产量，建立人用狂犬病疫苗（地鼠肾细胞）连续培养多次收获工艺。方法选用12～14日龄SPF地鼠，无菌取肾经消化，制备成细胞悬液，分装到40层细胞工厂并培养细胞成单层；接种狂犬病病毒固定毒aG株，连续培养病毒并多次收获。分别对同一细胞批制备的多个单次病毒收获液的免疫原性、病毒滴度和地鼠肾细胞蛋白质含量进行检测。结果用40层细胞工厂培养原代地鼠肾细胞和狂犬病病毒，细胞接种浓度为1．0×106～1．5×106cells /mL，（36±1）℃培养72 h成致密单层；按0．1 MOI病毒接种，可进行6次收获病毒；多个单次病毒收获液病毒滴度均不低于6．0 lgLD50/mL；免疫原性检查保护指数不低于100；地鼠肾细胞蛋白质残留量随着收获次数的增加而不断降低。结论用细胞工厂建立了人用狂犬病疫苗连续培养多次收获工艺，能显著提高地鼠肾单产量，增加产能。
目的：為提高生產效率、增加原代地鼠腎細胞單產量及狂犬病病毒產量，建立人用狂犬病疫苗（地鼠腎細胞）連續培養多次收穫工藝。方法選用12～14日齡SPF地鼠，無菌取腎經消化，製備成細胞懸液，分裝到40層細胞工廠併培養細胞成單層；接種狂犬病病毒固定毒aG株，連續培養病毒併多次收穫。分彆對同一細胞批製備的多箇單次病毒收穫液的免疫原性、病毒滴度和地鼠腎細胞蛋白質含量進行檢測。結果用40層細胞工廠培養原代地鼠腎細胞和狂犬病病毒，細胞接種濃度為1．0×106～1．5×106cells /mL，（36±1）℃培養72 h成緻密單層；按0．1 MOI病毒接種，可進行6次收穫病毒；多箇單次病毒收穫液病毒滴度均不低于6．0 lgLD50/mL；免疫原性檢查保護指數不低于100；地鼠腎細胞蛋白質殘留量隨著收穫次數的增加而不斷降低。結論用細胞工廠建立瞭人用狂犬病疫苗連續培養多次收穫工藝，能顯著提高地鼠腎單產量，增加產能。
목적：위제고생산효솔、증가원대지서신세포단산량급광견병병독산량，건립인용광견병역묘（지서신세포）련속배양다차수획공예。방법선용12～14일령SPF지서，무균취신경소화，제비성세포현액，분장도40층세포공엄병배양세포성단층；접충광견병병독고정독aG주，련속배양병독병다차수획。분별대동일세포비제비적다개단차병독수획액적면역원성、병독적도화지서신세포단백질함량진행검측。결과용40층세포공엄배양원대지서신세포화광견병병독，세포접충농도위1．0×106～1．5×106cells /mL，（36±1）℃배양72 h성치밀단층；안0．1 MOI병독접충，가진행6차수획병독；다개단차병독수획액병독적도균불저우6．0 lgLD50/mL；면역원성검사보호지수불저우100；지서신세포단백질잔류량수착수획차수적증가이불단강저。결론용세포공엄건립료인용광견병역묘련속배양다차수획공예，능현저제고지서신단산량，증가산능。
Objective To improve the production efficiency and increase primary hamster kidney cells ( PHKC) and rabies virus yield.To establish a new continuous culture and multiple harvests process using cell factory for rabies vaccine ( Ham-ster kidney cells ) for human use .Methods PHKC suspensions were prepared by asepsis digesting 12 to 14 day old pri-mary hamster kidney .The cells suspension was packed into cell factory and cultured to cell monolayer .Rabies fixed strains aG was inoculated and cultured ,and then multiple harvests were taken .Immunogenicity test , virus titer test and PHKC matrix proteins tset were taken for every single virus harvest from one cell batch .Result Cells cultures were carried out in CellSTACK?-40(40-layers cell factory) with cell density of 1.0×106-1.5×106cells /mL at (36±1) ℃ for 72 h.Cells were infected with rabies aG strains at 0.1 MOI and six virus harvests were manufactured from one cell batch .The titer of single virus harvests was not less than 6.0lgLD50/mL,and immunogenicity protection index was not less than 100.Cell ma-trix protein value reduced with the increase of virus harvest times .Conclusion The continuous culture and multiple har-vests process using cell factory for rabies vaccine ( primary hamster kidney cells ) for human use was established .It im-proved the production efficiency and increased PHKC and rabies virus yield .
