Cryovialzikoreshwa cyane mububiko bwa cryogenic bwo kubika imirongo ya selile nibindi bikoresho bikomeye bya biologiya, muri dewars yuzuye azote yuzuye.

Hariho ibyiciro byinshi bigira uruhare mukubungabunga neza selile muri azote yuzuye. Mugihe ihame ryibanze ari ugukonja buhoro, tekinike nyayo ikoreshwa biterwa nubwoko bwakagari na cryoprotectant yakoreshejwe. Hano haribintu byinshi byumutekano hamwe nibikorwa byiza ugomba kuzirikana mugihe ubitse selile kubushyuhe buke.

Iyi nyandiko igamije gutanga incamake yukuntu cryovial zibikwa muri azote yuzuye.

Cryovial ni iki

Cryovial ni ntoya, ifatanyirijwe hamwe igenewe kubika urugero rwamazi mubushyuhe buke cyane. Bemeza ko selile zabitswe muri cryoprotectant zidahura neza na azote yuzuye, bikagabanya ibyago byo kuvunika selile mugihe bikigirira akamaro gakonje gakabije ka azote.

Ububiko busanzwe buraboneka murwego rwubunini nubushushanyo - birashobora kuba imbere cyangwa hanze bifatanyirijwe hamwe nibisate binini cyangwa bizengurutse. Imiterere ya Sterile na sterile nayo irahari.

Ninde UkoreshaCyrovialkubika Ingirabuzimafatizo muri Azote

Urwego rwa NHS na laboratoire zigenga, hamwe n’ibigo by’ubushakashatsi bizobereye mu mabanki y’amaraso, epiteleial selile biologiya, immunologiya na biologiya selile ikoresha cryovial to selile cryopreserve.

Ingirabuzimafatizo zabitswe muri ubu buryo zirimo Utugari B na T, Utugari twa CHO, Hematopoietic Stem na Progenitor, Hybridoma, Uturemangingo two mu mara, Macrophage, Mesenchymal Stem na Progenitor, Monocytes, Myeloma, selile NK na Pluripotent Stem Cells.

Incamake yuburyo bwo kubika Cryovial muri Azote ya Liquid

Cryopreservation ni inzira ibika ingirabuzimafatizo hamwe n’ibindi binyabuzima bikonjesha mu bushyuhe buke cyane. Ingirabuzimafatizo zirashobora kubikwa muri azote yuzuye mumyaka nta gutakaza imbaraga za selile. Uru ni urucacagu rw'uburyo bukoreshwa.

Gutegura Akagari

Uburyo nyabwo bwo gutegura ingero buratandukana bitewe nubwoko bwakagari, ariko muri rusange, ingirabuzimafatizo zegeranijwe kandi zegeranijwe kugirango zitezimbere pellet ikungahaye. Iyi pellet noneho ihagarikwa muri supernatant ivanze na cryoprotectant cyangwa uburyo bwo kubika ibintu.

Kurinda hagati

Ubu buryo bukoreshwa mu kubungabunga ingirabuzimafatizo ziri mu bushyuhe buke bazakorerwa babuza kwishyiriraho kristu zo mu nda ndetse n’inyuma zidasanzwe bityo urupfu rw ingirabuzimafatizo. Uruhare rwabo ni ugutanga ahantu hizewe, harinda ingirabuzimafatizo hamwe nuduce mugihe cyo gukonjesha, kubika, no gusya.

Igikoresho nka plasma nshya ikonje (FFP), igisubizo cya heparinised plasmalyte cyangwa serumu idafite, ibisubizo bitarimo inyamanswa bivangwa na cryoprotectants nka dimethyl sulphoxide (DMSO) cyangwa glycerol.

Icyitegererezo cya pellet yongeye gusukwa muri polypropilene cryovial nkaSuzhou Ace Biomedical societe Cryogenic Ububiko.

Ni ngombwa kutuzuza cryovial kuko ibi bizongera ibyago byo guturika no gusohora ibirimo (1).

Igipimo cyo gukonjesha

Muri rusange, igipimo cyogukonjesha gahoro gahoro gikoreshwa muburyo bwiza bwo kubika ingirabuzimafatizo.

Ingero zimaze guhindurwa mumashanyarazi, zishyirwa kurubura rutose cyangwa muri firigo ya 4 and hanyuma uburyo bwo gukonjesha butangira muminota 5. Nkuyobora muri rusange, selile zikonjeshwa ku gipimo cya -1 kugeza kuri 3 ku munota (2). Ibi bigerwaho hifashishijwe icyuma gikonjesha cyangwa ugashyira ibibindi mumasanduku yiziritse yashyizwe muri 70 ° C kugeza kuri 90 ° C.

Kwimura Azote

Ibikonjo byafunzwe bya kirogenike noneho byimurirwa mumazi ya azote yuzuye mugihe kitazwi mugihe ubushyuhe buri munsi ya -135 ℃ bugumaho.

Ubu bushyuhe buke cyane burashobora kuboneka mukwibiza mumazi ya azote.

Icyiciro cyamazi cyangwa imyuka?

Kubika icyiciro cya azote azwiho kugumana ubushyuhe bukonje kandi buhoraho, ariko akenshi ntibisabwa kubwimpamvu zikurikira:

• Gukenera ubunini bunini (ubujyakuzimu) bwa azote yuzuye ishobora guteza akaga. Gutwika cyangwa guhumeka bitewe nibi byago rwose.
• Ibibazo byanduye byanduye byanduye nka aspergillus, hep B na virusi bikwirakwizwa hakoreshejwe amazi ya azote (2,3)
• Ubushobozi bwa azote yamazi yatemba mumibindi mugihe cyo kwibiza. Iyo ikuwe mububiko igashyuha ubushyuhe bwicyumba, azote iraguka vuba. Kubera iyo mpamvu, isahani irashobora kumeneka iyo ikuwe mububiko bwa azote yuzuye, bigatera akaga imyanda iguruka no guhura nibirimo (1, 4).

Kubera izo mpamvu, kubika ubushyuhe bukabije cyane ni muri azote. Mugihe ibyitegererezo bigomba kubikwa mugice cyamazi, hagomba gukoreshwa igituba cyihariye cya cryoflex.

Ikibi cyumuyaga ni uko ubushyuhe bwikigereranyo bushobora kugaragara bigatuma ihindagurika ryubushyuhe hagati ya -135 ℃ na -190 ℃. Ibi birasaba gukurikiranira hafi kandi ushishikaye urugero rwa azote yuzuye hamwe nubushyuhe butandukanye (5).

Ababikora benshi basaba ko cryovial ikwiriye kubikwa kugeza kuri -135 ℃ cyangwa gukoreshwa mugice cyumuyaga gusa.

Gukurura Utugingo twawe twirinda

Uburyo bwo gusya burahangayikishije umuco wafunzwe, kandi harasabwa gufata neza nubuhanga kugirango habeho kubaho neza, gukira, nimikorere ya selile. Porotokole yukuri izaterwa nubwoko bwakagari. Ariko, gusya byihuse bifatwa nkibisanzwe kuri:

• Kugabanya ingaruka zose zo kugarura selile
• Fasha kugabanya igihe cyo kwerekana ibisubizo biboneka mubitangazamakuru bikonje
• Mugabanye ibyangiritse byose hamwe na ice rerystallisation

Ubwogero bwamazi, ubwogero bwamasaro, cyangwa ibikoresho byabigenewe byifashishwa mugukata ingero.

Kenshi na kenshi umurongo 1 w'akagari ushwanyagujwe icyarimwe muminota 1-2, ukizunguruka witonze mu bwogero bwa 37 until kugeza igihe hasigaye urubura ruto gusa muri vial mbere yuko rwozwa muburyo bwo gukura mbere.

Kuri selile zimwe nkinsoro zinyamabere, gushyuha gahoro ningirakamaro kugirango babeho.

Ingirabuzimafatizo ubu ziteguye umuco w’akagari, kwigunga kw’akagari, cyangwa ku bijyanye n’ingirabuzimafatizo ya haematopoietic - ubushakashatsi bufatika bwo kwemeza ubusugire bw’utugingo ngengabuzima mbere yo kuvura myeloablative.

Nibisanzwe gufata alikoti ntoya yicyitegererezo cyakoreshejwe mukubara selile kugirango hamenyekane utugingo ngengabuzima dushyira mu muco. Urashobora noneho gusuzuma ibisubizo byuburyo bwo gutandukanya akagari hanyuma ukamenya selile selile.

Imyitozo myiza yo kubika Cryovial

Uburyo bwiza bwo kubika ibyitegererezo bibitswe muri cryovial biterwa nibintu byinshi muri protocole harimo kubika neza no kubika inyandiko.

• Gabanya selile hagati yububiko- Niba inomero zibyemerera, gabanya selile hagati ya vial hanyuma ubibike ahantu hatandukanye kugirango ugabanye ibyago byo gutakaza icyitegererezo kubera kunanirwa ibikoresho.
• Irinde kwanduzanya- Hitamo uburyo bumwe bwo gukoresha sterile cryogenic vial cyangwa autoclave mbere yo gukoreshwa nyuma
• Koresha inzabya nini zikwiye kuri selile yawe- ibibindi biza mubice byinshi hagati ya 1 na 5mls. Irinde kuzuza ibibindi byinshi kugirango ugabanye ibyago byo guturika.
• Hitamo imbere cyangwa hanze yinyuma ya cryogenic- Imbere yimbere yimbere irasabwa na kaminuza zimwe murwego rwo kubungabunga umutekano - zirashobora kandi kwirinda kwanduza mugihe cyo kuzuza cyangwa iyo zibitswe muri azote yuzuye.
• Irinde kumeneka- Koresha kashe yatewe inshinge zibumbwe muri screw-cap cyangwa O-impeta kugirango wirinde kumeneka no kwanduza.
• Koresha 2D barcode hamwe na label vial- kugirango ukurikirane neza, ibibindi bifite ahantu hanini ho kwandika bituma buri vial iba yanditseho bihagije. 2D barcode irashobora gufasha mubuyobozi bwo kubika no kubika inyandiko. Ibara ryanditseho amabara ni ingirakamaro kubimenyekanisha byoroshye.
• Kubungabunga bihagije- Kugirango selile zidatakara, ibikoresho byo kubika bigomba guhora bikurikirana ubushyuhe nurwego rwa azote. Imenyekanisha rigomba gushyirwaho kugirango umenyeshe abakoresha amakosa.

Kwirinda Umutekano

Azote y'amazi imaze kuba akamenyero mubushakashatsi bugezweho ariko itwara ibyago byo gukomeretsa bikomeye iyo ikoreshejwe nabi.

Ibikoresho bikingira umuntu ku giti cye (PPE) bigomba kwambarwa kugirango bigabanye ingaruka ziterwa nubukonje, gutwikwa nibindi bintu bibi mugihe ukoresha azote yuzuye. Kwambara

• Uturindantoki twa Cryogenic
• Ikoti rya laboratoire
• Ingaruka irwanya isura yuzuye yuzuye nayo itwikiriye ijosi
• Inkweto zifunze
• Amashanyarazi adafite amashanyarazi

Firigo ya azote yuzuye igomba gushyirwa ahantu hafite umwuka uhagije kugirango hagabanuke ibyago byo guhumeka - azote ihumeka kandi ikuraho ogisijeni yo mu kirere. Ububiko bunini bugomba kugira sisitemu yo gutabaza ya ogisijeni.

Gukorera muri babiri mugihe ukoresha azote yuzuye nibyiza kandi kuyikoresha hanze yamasaha asanzwe yakazi bigomba kubuzwa.

Cryovial kugirango ushyigikire akazi kawe

Isosiyete ya Suzhou Ace Biomedical itanga ibicuruzwa byinshi byujuje ibyifuzo byawe byo kubika ibintu bitandukanye byingirabuzimafatizo. Inshingano zirimo urutonde rwa tubes kimwe nurwego rwa sterile cryovial.

Cryovial yacu ni:

• Ibikoresho bya Laboratwari 0.5mL 1.5mL 2.0mL Cryovial Cryogenic Vial Conical Hasi Cryotube hamwe na Gasket

  ● 0.5ml, 1.5ml, 2.0ml ibisobanuro, hamwe nijipo cyangwa idafite ijipo
  Design Igishushanyo mbonera cyangwa kwihagararaho, sterile cyangwa non-sterile byombi birahari
  Imiyoboro ya kaburimbo ikozwe mubyiciro byubuvuzi polypropilene
  ● PP Cryotube Vial irashobora gukonjeshwa inshuro nyinshi kandi igashonga
  Igishushanyo mbonera cyo hanze gishobora kugabanya amahirwe yo kwanduza mugihe cyo kuvura icyitegererezo.
  Shira cap cap cryogenic tubes Universal screw umugozi wo gukoresha
  Ibijumba bikwiranye na rotor nyinshi
  ● Cryogenic tube o-ring tubes ihuye na santimetero 1 na santimetero 2, 48well, 81well ， 96wiza na 100well frigo
  ● Autoclavable to 121 ° C kandi irashobora gukonja kuri -86 ° C.

  IGICE CYA OYA	IMIKORESHEREZE	UMUBUMBE	CAPAMABARA	PCS /BAG	URUBUGA / URUBANZA
  ACT05-BL-N	PP	0.5ML	Umukara, Umuhondo, Ubururu, Umutuku, Umutuku, Umweru	500	10
  ACT15-BL-N	PP	1.5ML	Umukara, Umuhondo, Ubururu, Umutuku, Umutuku, Umweru	500	10
  ACT15-BL-NW	PP	1.5ML	Umukara, Umuhondo, Ubururu, Umutuku, Umutuku, Umweru	500	10
  ACT20-BL-N	PP	2.0ML	Umukara, Umuhondo, Ubururu, Umutuku, Umutuku, Umweru	500	10