Kimwe mu byiza byingenzi byumuco wutugari nubushobozi bwo gukoresha chimie yumubiri yimyororokere (ni ukuvuga ubushyuhe, pH, umuvuduko wa osmotic, impagarara za O2 na CO2) hamwe nibidukikije byumubiri (ni ukuvuga imisemburo hamwe nintungamubiri).Usibye ubushyuhe, ibidukikije byumuco bigenzurwa nuburyo bwo gukura.
Nubwo ibidukikije byimiterere yumuco bidasobanutse nkibidukikije byumubiri nubumara, gusobanukirwa neza nibigize serumu, kumenya ibintu bikura bikenerwa kugirango ikwirakwizwa, hamwe no gusobanukirwa neza ibidukikije byingirabuzimafatizo mumuco..
1.Ibidukikije byumuco bigira ingaruka kumikurire
Nyamuneka menya ko imiterere yumuco utugari itandukanye kuri buri bwoko bwakagari.
Ingaruka zo gutandukira kumiterere yumuco isabwa kubwoko bwakagari runaka uhereye kumagambo ya fenotipike idasanzwe kugeza kunanirwa burundu kumuco.Kubwibyo, turagusaba ko umenyera umurongo wa selire ushimishijwe kandi ugakurikiza byimazeyo amabwiriza yatanzwe kuri buri gicuruzwa ukoresha mubushakashatsi bwawe.
2.Ibyitonderwa byo gushyiraho uburyo bwiza bwimikorere yumuco wa selile:
Itangazamakuru ryumuco na serumu (reba hano hepfo kubindi bisobanuro)
urwego pH na CO2 (reba hano hepfo kubindi bisobanuro)
Guhinga plastike (reba hano hepfo kubindi bisobanuro)
Ubushyuhe (reba hano hepfo kubindi bisobanuro)
2.1 Itangazamakuru ryumuco na Serumu
Umuco uciriritse nigice cyingenzi cyibidukikije byumuco, kuko utanga intungamubiri, ibintu bikura hamwe na hormone zikenerwa mugukura kwakagari, kandi bikagenga umuvuduko wa pH na osmotic wumuco.
Nubwo ubushakashatsi bwambere bwumuco bwakagari bwakozwe hifashishijwe itangazamakuru risanzwe ryakuwe mubikomoka ku ngingo hamwe n’amazi yo mu mubiri, gukenera ubuziranenge, ubwiza bw’itangazamakuru, no gukenera kwiyongera byatumye habaho itangazamakuru risobanutse.Ubwoko butatu bwibanze bwitangazamakuru ni itangazamakuru ryibanze, kugabanya itangazamakuru rya serumu nibitangazamakuru bidafite serumu, kandi bifite ibisabwa bitandukanye kugirango hongerwe serumu.
2.1.1
Umuco w'akagari ka Gibco
Imirongo myinshi ya selile ikura neza mubitangazamakuru byibanze birimo aside amine, vitamine, imyunyu ngugu, hamwe na karubone (nka glucose), ariko ibyo bitangazamakuru byibanze bigomba kongerwaho serumu.
2.1.2 Kugabanya serumu ikoreshwa
Icupa hamwe na Gibco Serumu Hagati
Iyindi ngamba yo kugabanya ingaruka mbi za serumu mubigeragezo byumuco w'akagari ni ugukoresha itangazamakuru ryagabanijwe.Kugabanya serumu ikoreshwa ni formulaire yibanze ikungahaye ku ntungamubiri nibintu bikomoka ku nyamaswa, bishobora kugabanya urugero rwa serumu isabwa.
2.1.3
Icupa hamwe na Gibco serumu idafite
Uburyo butagira serumu (SFM) buzenguruka ikoreshwa rya serumu yinyamanswa mugusimbuza serumu nimirire ikwiye hamwe na hormone.Imico myinshi yibanze n'imirongo y'utugari bifite serumu idafite serumu, harimo umushinwa Hamster Ovary (CHO) recombinant protein itanga umurongo, imirongo itandukanye ya Hybridoma, imirongo y'udukoko Sf9 na Sf21 (Spodoptera frugiperda), ndetse no kubakira virusi. (kurugero, 293, VERO, MDCK, MDBK), nibindi.Imbonerahamwe ikurikira irerekana ibyiza nibibi byitangazamakuru ridafite serumu.
Ibyiza
Ongera usobanutse
Imikorere ihamye
Korozwa byoroshye no gutunganya epfo na ruguru
Suzuma neza imikorere ya selile
Ongera umusaruro
Kugenzura neza imyitwarire yumubiri
Kumenyekanisha itangazamakuru rya selile
Ingaruka
Ubwoko bwakagari bwihariye bwihariye busabwa
Ukeneye kwera kwinshi
Gutinda gukura
2.2.1 pH urwego
Imirongo myinshi isanzwe yinyamabere ikura neza kuri pH 7.4, kandi itandukaniro riri hagati yimirongo itandukanye ni nto.Nyamara, imirongo imwe nimwe yahinduwe yerekanwe gukura neza mubidukikije bya acide nkeya (pH 7.0 - 7.4), mugihe imirongo imwe isanzwe ya fibroblast ihitamo ibidukikije bya alkaline nkeya (pH 7.4 - 7.7).Imirongo y'udukoko nka Sf9 na Sf21 ikura neza kuri pH 6.2.
Urwego rwa CO2
Uburyo bwo gukura bugenzura pH yumuco kandi bugahindura ingirabuzimafatizo mumuco kugirango zirwanye impinduka muri pH.Mubisanzwe, iyi buffering igerwaho harimo organic (urugero, HEPES) cyangwa CO2-bicarbonate ishingiye kuri buffer.Kuberako pH yikigereranyo iterwa nuburinganire bworoshye bwa dioxyde de carbone yashonze (CO2) na bicarbonate (HCO3-), impinduka za CO2 zo mu kirere zizahindura pH yikigereranyo.Kubwibyo, mugihe ukoresheje icyuma giciriritse hamwe na CO2-bicarbonate ishingiye kuri buffer, birakenewe gukoresha CO2 idasanzwe, cyane cyane iyo utera utugingo ngengabuzima mu mafunguro y’umuco ufunguye cyangwa ugahindura imirongo ya selile ihindagurika cyane.Nubwo abashakashatsi benshi bakoresha CO2 5-7% mukirere, ubushakashatsi bwinshi bwumuco bwakagari bukoresha 4-10% CO2.Nyamara, buri cyiciro gifite ibyifuzo bya CO2 hamwe nubushakashatsi bwa bicarbonate kugirango ugere kumuvuduko mwiza wa pH na osmotic;kubindi bisobanuro, nyamuneka reba amabwiriza yakozwe n'abaciriritse.
2.3 Guhinga plastike
Plastiki yumuco utugari iraboneka muburyo butandukanye, ingano nubuso kugirango bikwiranye numuco utandukanye wimikorere.Koresha umuco wumudugudu wa plastike yubuso hamwe nubuyobozi bwumuco wibikoresho munsi kugirango bigufashe guhitamo plastike ibereye yo gukoresha umuco wawe.
Reba ibintu byose bya Thermo Scientific Nunc selile yumuco plastike (ihuza ryamamaza)
2.4 Ubushyuhe
Ubushyuhe bwiza bwumuco utugari biterwa ahanini nubushyuhe bwumubiri wa nyirarureshwa uturemangingo twitaruye, kandi ku rugero ruto ku mpinduka zidasanzwe mu bushyuhe (urugero, ubushyuhe bwuruhu bushobora kuba munsi yubwa imitsi ya skeletale ).Ku muco w'akagari, Ubushyuhe ni ikibazo gikomeye kuruta gushyuha.Kubwibyo, ubushyuhe muri incubator busanzwe bushyirwa munsi yubushyuhe bwiza.
2.4.1 Ubushyuhe bwiza kumirongo itandukanye
Imirongo myinshi yumuntu ninyamabere ibikwa kuri 36 ° C kugeza 37 ° C kugirango ikure neza.
Ingirabuzimafatizo zihingwa kuri 27 ° C kugirango zikure neza;zikura buhoro buhoro ku bushyuhe bwo hasi n'ubushyuhe buri hagati ya 27 ° C na 30 ° C.Hejuru ya 30 ° C, imbaraga za selile zudukoko ziragabanuka, niyo yagaruka kuri 27 ° C, selile ntizakira.
Imirongo ya selile ikenera 38.5 ° C kugirango igere ku mikurire nini.Nubwo izo selile zishobora kubikwa kuri 37 ° C, zizakura buhoro buhoro.
Imirongo ya selile ikomoka ku nyamaswa zifite amaraso akonje (nka amphibian, amafi yo mu mazi akonje) irashobora kwihanganira ubushyuhe bugari bwa 15 ° C kugeza 26 ° C.
Igihe cyo kohereza: Gashyantare-01-2023