| Área: Virologia ( Divisão P ) OPTIMIZATION OF THE ROTAVIRUS PRODUCTION USING BIOREACTOR Rosana de Lima Paoli (IBU); Flávia Grecco Giacomin (IBU); Luiz Fernando de Oliveira Gomes (IBU); Neuza Maria Frazatti Gallina (IBU)
ResumoIntroduction: Rotavirus disease is associated with 600.000 deaths worldwide among children younger than five years age. Most symptomatic episodes occur in young children between the ages of 3 months and 2 years. The development of rotavirus vaccine is very important to prevent the severe rotavirus disease. This vaccine is prepared with rotavirus suspensions obtained of cell cultures infected with different rotavirus serotypes. 㰀倀 猀琀礀氀攀㴀∀吀䔀堀吀ⴀ䄀䰀䤀䜀一㨀 樀甀猀琀椀昀礀㬀 䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䴀䄀刀䜀䤀一㨀 挀洀 挀洀 瀀琀∀ 挀氀愀猀猀㴀䴀猀漀一漀爀洀愀氀㸀㰀匀倀䄀一 猀琀礀氀攀㴀∀䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䘀伀一吀ⴀ䘀䄀䴀䤀䰀夀㨀 ✀吀椀洀攀猀 一攀眀 刀漀洀愀渀✀Ⰰ✀猀攀爀椀昀✀㬀 䌀伀䰀伀刀㨀 戀氀愀挀欀㬀 䘀伀一吀ⴀ匀䤀娀䔀㨀 ㈀瀀琀∀ 氀愀渀最㴀䔀一ⴀ唀匀㸀伀戀樀攀挀琀椀瘀攀猀㨀 吀栀攀 漀戀樀攀挀琀椀瘀攀 漀昀 琀栀椀猀 猀琀甀搀礀 眀愀猀 琀㰀⼀匀倀䄀一㸀㰀匀倀䄀一 猀琀礀氀攀㴀∀䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䘀伀一吀ⴀ䘀䄀䴀䤀䰀夀㨀 ✀吀椀洀攀猀 一攀眀 刀漀洀愀渀✀Ⰰ✀猀攀爀椀昀✀㬀 䘀伀一吀ⴀ匀䤀娀䔀㨀 ㈀瀀琀∀ 氀愀渀最㴀䔀一ⴀ唀匀㸀漀 攀瘀愀氀甀愀琀攀 琀栀攀 瀀漀琀攀渀挀礀 爀漀琀愀瘀椀爀甀猀 猀甀猀瀀攀渀猀椀漀渀 瀀爀漀搀甀挀攀搀 椀渀 戀椀漀爀攀愀挀琀漀爀 ⠀氀愀爀最攀 猀挀愀氀攀⤀ 琀栀攀 猀攀爀漀琀礀瀀攀猀 㰀匀倀䄀一 挀氀愀猀猀㴀猀琀礀氀攀 㸀㰀匀倀䄀一 猀琀礀氀攀㴀∀䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䘀伀一吀ⴀ䘀䄀䴀䤀䰀夀㨀 ✀吀椀洀攀猀 一攀眀 刀漀洀愀渀✀Ⰰ✀猀攀爀椀昀✀㬀 洀猀漀ⴀ愀渀猀椀ⴀ昀漀渀琀ⴀ猀椀稀攀㨀 ㈀⸀ 瀀琀㬀 洀猀漀ⴀ戀椀搀椀ⴀ昀漀渀琀ⴀ猀椀稀攀㨀 ㈀⸀ 瀀琀∀㸀䜀Ⰰ 䜀㈀Ⰰ 䜀㌀Ⰰ 䜀㐀 愀渀搀 䜀㤀⸀㰀⼀匀倀䄀一㸀㰀⼀匀倀䄀一㸀㰀䈀 猀琀礀氀攀㴀∀洀猀漀ⴀ戀椀搀椀ⴀ昀漀渀琀ⴀ眀攀椀最栀琀㨀 渀漀爀洀愀氀∀㸀㰀漀㨀瀀㸀㰀⼀漀㨀瀀㸀㰀⼀䈀㸀㰀⼀匀倀䄀一㸀㰀⼀倀㸀
Methods and Materials: Vero cells added on solid microcarriers (cytodex 1) were cultivated with serum free- medium in bioreactor of 5L (Celligem NBS) with initial concentration cellular of 1, 0 x 109 cells. After three or four days (100% cell confluence) the cells were infected with different rotavirus serotypes (reassortant strains G1, G2, G3, G4 and G9 from NIH). Samples this supernatant was taken for observed cytopatic effects and cellular concentration. When observed 90-100% of cytopatic effects, the harvests of the supernatant of these cultures were realized and the samples were taken to determine the rotavirus titers by PFU (Plate Forming Units) test. For this test were utilized MA-104 cells cultivated in plate six wells. 㰀倀 猀琀礀氀攀㴀∀吀䔀堀吀ⴀ䄀䰀䤀䜀一㨀 樀甀猀琀椀昀礀㬀 䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䴀䄀刀䜀䤀一㨀 挀洀 挀洀 瀀琀∀ 挀氀愀猀猀㴀䴀猀漀一漀爀洀愀氀㸀㰀匀倀䄀一 猀琀礀氀攀㴀∀䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䘀伀一吀ⴀ䘀䄀䴀䤀䰀夀㨀 ✀吀椀洀攀猀 一攀眀 刀漀洀愀渀✀Ⰰ✀猀攀爀椀昀✀㬀 䘀伀一吀ⴀ匀䤀娀䔀㨀 ㈀瀀琀∀ 氀愀渀最㴀䔀一ⴀ唀匀㸀刀攀猀甀氀琀猀㨀 吀栀攀 琀椀琀攀爀猀 漀戀琀愀椀渀攀搀 椀渀 琀栀攀 猀愀洀瀀氀攀猀 漀昀 琀栀攀 栀愀爀瘀攀猀琀 眀攀爀攀 㰀匀唀倀㸀㘀⸀㈀㰀⼀匀唀倀㸀Ⰰ 㰀匀唀倀㸀㘀⸀㌀㰀⼀匀唀倀㸀Ⰰ 㰀匀唀倀㸀㘀⸀㈀㰀⼀匀唀倀㸀Ⰰ 㰀匀唀倀㸀㘀⸀ 㰀⼀匀唀倀㸀 愀渀搀 㰀漀㨀瀀㸀㰀⼀漀㨀瀀㸀㰀⼀匀倀䄀一㸀㰀⼀倀㸀
107, 5 PFU/ml for G1, G2, G3, G4 and G9 rotavirus serotypes respectively. 㰀倀 猀琀礀氀攀㴀∀吀䔀堀吀ⴀ䄀䰀䤀䜀一㨀 樀甀猀琀椀昀礀㬀 䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䴀䄀刀䜀䤀一㨀 挀洀 挀洀 瀀琀∀ 挀氀愀猀猀㴀䴀猀漀一漀爀洀愀氀㸀㰀匀倀䄀一 猀琀礀氀攀㴀∀䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀 ─㬀 䘀伀一吀ⴀ䘀䄀䴀䤀䰀夀㨀 ✀吀椀洀攀猀 一攀眀 刀漀洀愀渀✀Ⰰ✀猀攀爀椀昀✀㬀 䘀伀一吀ⴀ匀䤀娀䔀㨀 ㈀瀀琀∀ 氀愀渀最㴀䔀一ⴀ唀匀㸀䐀椀猀挀甀猀猀椀漀渀㨀㰀䈀 猀琀礀氀攀㴀∀洀猀漀ⴀ戀椀搀椀ⴀ昀漀渀琀ⴀ眀攀椀最栀琀㨀 渀漀爀洀愀氀∀㸀 㰀⼀䈀㸀吀栀攀 琀椀琀攀爀猀 昀漀甀渀搀 椀渀 琀栀攀 猀甀瀀攀爀渀愀琀愀渀琀 漀昀 琀栀攀 挀甀氀琀甀爀攀猀 椀渀昀攀挀琀攀搀 眀椀琀栀 搀椀昀昀攀爀攀渀琀 爀漀琀愀瘀椀爀甀猀 猀攀爀漀琀礀瀀攀猀 猀栀漀眀攀搀 琀栀愀琀 琀栀攀 甀琀椀氀椀稀愀琀椀漀渀 漀昀 嘀攀爀漀 挀攀氀氀 愀搀搀攀搀 漀渀 猀漀氀椀搀 洀椀挀爀漀挀愀爀爀椀攀爀猀 挀甀氀琀椀瘀愀琀攀搀 椀渀 戀椀漀爀攀愀挀琀漀爀 椀猀 愀 最漀漀搀 猀琀爀愀琀攀最礀 琀漀 漀瀀琀椀洀椀稀攀 琀栀攀 爀漀琀愀瘀椀爀甀猀 瀀爀漀搀甀挀琀椀漀渀 愀琀 䤀渀猀琀椀琀甀琀攀 䈀甀琀愀渀琀愀渀⸀ 㰀漀㨀瀀㸀㰀⼀漀㨀瀀㸀㰀⼀匀倀䄀一㸀㰀⼀倀㸀㰀匀倀䄀一 猀琀礀氀攀㴀∀䰀䤀一䔀ⴀ䠀䔀䤀䜀䠀吀㨀 㔀─㬀 䘀伀一吀ⴀ䘀䄀䴀䤀䰀夀㨀 ✀吀椀洀攀猀 一攀眀 刀漀洀愀渀✀Ⰰ✀猀攀爀椀昀✀㬀 䘀伀一吀ⴀ匀䤀娀䔀㨀 ㈀瀀琀㬀 洀猀漀ⴀ昀愀爀攀愀猀琀ⴀ氀愀渀最甀愀最攀㨀 䔀一ⴀ唀匀㬀 洀猀漀ⴀ昀愀爀攀愀猀琀ⴀ昀漀渀琀ⴀ昀愀洀椀氀礀㨀 䌀愀氀椀戀爀椀㬀 洀猀漀ⴀ愀渀猀椀ⴀ氀愀渀最甀愀最攀㨀 䔀一ⴀ唀匀㬀 洀猀漀ⴀ戀椀搀椀ⴀ氀愀渀最甀愀最攀㨀 䄀刀ⴀ匀䄀∀ 氀愀渀最㴀䔀一ⴀ唀匀㸀䘀椀渀愀渀挀椀愀氀 猀甀瀀瀀漀爀琀㨀 䈀甀琀愀渀琀愀渀 䘀漀甀渀搀愀琀椀漀渀㰀⼀匀倀䄀一㸀㰀⼀昀漀渀琀㸀㰀⼀瀀㸀㰀戀爀㸀㰀戀㸀倀愀氀愀瘀爀愀猀ⴀ挀栀愀瘀攀㨀 㰀⼀戀㸀☀渀戀猀瀀㬀䈀椀漀爀攀愀挀琀漀爀Ⰰ 刀漀琀愀瘀椀爀甀猀Ⰰ 嘀愀挀挀椀渀攀㰀⼀琀搀㸀㰀⼀琀爀㸀㰀⼀琀愀戀氀攀㸀㰀⼀琀爀㸀㰀⼀琀搀㸀㰀⼀琀愀戀氀攀㸀㰀⼀戀漀搀礀㸀㰀⼀栀琀洀氀㸀
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