Ama-Cryovialszivame ukusetshenziselwa isitoreji cryogenic of cell lines kanye nezinye izinto ezibalulekile biological, dewar egcwele liquid nitrogen.

Kunezigaba eziningana ezihilelekile ekulondolozweni ngempumelelo kwamaseli ku-nitrogen ewuketshezi. Nakuba umgomo oyisisekelo uwukuqandisa okunensayo, indlela okuyiyonayona esetshenziswayo incike ohlotsheni lweseli kanye ne-cryoprotectant esetshenzisiwe. Kukhona ukucatshangelwa okuningana kokuphepha kanye nezinqubo ezingcono kakhulu okufanele zicatshangelwe lapho ugcina amaseli kumazinga okushisa aphansi kangaka.

Lokhu okuthunyelwe kuhlose ukunikeza umbono jikelele wokuthi ama-cryovials agcinwa kanjani ku-nitrogen ewuketshezi.

Yini ama-Cryovials

Ama-Cryovials amabhodlela amancane, anekepisi aklanyelwe ukugcina amasampula oketshezi emazingeni okushisa aphansi kakhulu. Baqinisekisa ukuthi amaseli alondolozwe ku-cryoprotectant awathintani ngokuqondile ne-nitrogen ewuketshezi, okunciphisa ubungozi bokuphuka kwamaselula ngesikhathi esazuza kumphumela wokupholisa owedlulele we-nitrogen ewuketshezi.

Izitsha zivame ukutholakala ezinhlobonhlobo zemiqulu kanye nemiklamo - zingafakwa ngaphakathi noma ngaphandle ngemicu ephansi noma eyindilinga. Amafomethi ayinyumba nangawona ayatholakala.

Ubani OsebenzisaAma-CyrovialsUkugcina Amaseli Ku-Nitrogen Ewuketshezi

Uhlu lwama-NHS namalabhorethri azimele, kanye nezikhungo zocwaningo ezigxile ekugcinweni kwegazi ngezintambo, i-epithelial cell biology, i-immunology kanye ne-stem cell biology zisebenzisa ama-cryovials ukuze kugcinwe amaseli.

Amangqamuzana alondolozwe ngale ndlela ahlanganisa Amangqamuzana e-B no-T, ama-CHO Cells, i-Hematopoietic Stem kanye nama-Progenitor Cells, ama-Hybridomas, ama-Intestinal Cells, ama-Macrophages, ama-Mesenchymal Stem kanye nama-Progenitor Cells, ama-Monocytes, i-Myeloma, i-NK Cells kanye nama-Pluripotent Stem Cells.

Uhlolojikelele Lwendlela Yokugcina I-Cryovials ku-Nitrogen Yamanzi

I-Cryopreservation yinqubo egcina amaseli nezinye izinto eziphilayo ngokuzipholisa emazingeni okushisa aphansi kakhulu. Amaseli angagcinwa ku-nitrogen ewuketshezi iminyaka ngaphandle kokulahlekelwa ukusebenza kwamaseli. Lolu wuhlaka lwezinqubo ezisetshenziswayo.

Ukulungiselela Iseli

Indlela eqondile yokulungiselela amasampula izohluka kuye ngohlobo lweseli, kodwa ngokuvamile, amaseli ayaqoqwa futhi afakwe phakathi ukuze akhe i-pellet enothe ngeseli. Le pellet ibe isiphinde yamiswa ku-supernatant exutshwe ne-cryoprotectant noma i-cryopreservation medium.

I-Cryopreservation Medium

Le ndlela yokuxhumana isetshenziselwa ukulondoloza amaseli ezindaweni ezinezinga lokushisa eliphansi azobhekana nazo ngokuvimbela ukwakheka kwamakristalu angaphakathi nangaphandle kwamangqamuzana futhi ngenxa yalokho ukufa kwamangqamuzana. Indima yabo iwukunikeza indawo ephephile, evikelayo yamangqamuzana nezicubu ngesikhathi sokuqandisa, ukugcinwa, kanye nezinqubo zokuncibilika.

I-medium efana ne-fresh frozen plasma (FFP), isixazululo se-plasmalyte esine-heparinised noma i-serum-free, izixazululo ezingenayo ingxenye yezilwane zixutshwa nama-cryoprotectants afana ne-dimethyl sulphoxide (DMSO) noma i-glycerol.

I-pellet yesampula eyenziwe kabusha i-aliquated ibe yi-polypropylene cryovials njengeInkampani yeSuzhou Ace Biomedical Cryogenic Storage Vials.

Kubalulekile ukuthi ungagcwalisi ngokweqile ama-cryovials njengoba lokhu kuzonyusa ingozi yokuqhekeka kanye nokukhishwa okungenzeka kokuqukethwe (1).

Izinga Lokuqanda Elilawulwayo

Ngokuvamile, izinga lokuqandisa elilawulwa kancane lisetshenziselwa ukugcinwa ngempumelelo kwamaseli.

Ngemuva kokuthi amasampula efakwe ezitsheni ze-cryogenic, abekwa eqhweni elimanzi noma esiqandisini esingu-4℃ futhi inqubo yokuqandisa iqalwa phakathi nemizuzu emi-5. Njengomhlahlandlela ojwayelekile, amaseli apholiswa ngesilinganiso -1 kuya ku -3 ngomzuzu (2). Lokhu kufezwa kusetshenziswa isibandisi esipholile esihlelekayo noma ngokubeka izitsha ebhokisini elifakwe intuba elibekwe ku--70°C kuya ku-90°C efrijini elilawulwayo.

Dlulisela ku-Liquid Nitrogen

Izitsha eziqandisiwe ze-cryogenic zibe sezidluliselwa ethangini le-nitrogen eliwuketshezi isikhathi esingaziwa inqobo nje uma izinga lokushisa elingaphansi kuka -135℃ ligcinwa.

Lawa mazinga okushisa aphansi kakhulu angatholwa ngokucwiliswa ku-nitrogen yesigaba soketshezi noma somhwamuko.

I-Liquid noma i-Vapor Phase?

Ukugcinwa kwe-nitrogen yesigaba esiwuketshezi kwaziwa ngokugcina izinga lokushisa elibandayo ngokungaguquguquki ngokuphelele, kodwa ngokuvamile akunconywa ngenxa yezizathu ezilandelayo:

• Isidingo semithamo emikhulu (ukujula) kwe-nitrogen engamanzi okungaba yingozi. Ukushiswa noma ukuqunjelwa ngenxa yalokhu kuyingozi yangempela.
• Izigameko ezibhalwe phansi zokuthelelana ngezifo ezithathelwanayo ezifana ne-aspergillus, i-hep B nokusabalala kwegciwane nge-liquid nitrogen medium (2,3)
• Amathuba okuthi initrogen eyiketshezi ivuze ezitsheni ngesikhathi sokucwiliswa. Lapho isuswa endaweni yokugcina futhi ifudunyezwa ekamelweni lokushisa, i-nitrogen ikhula ngokushesha. Ngenxa yalokho, isitsha singaphahlazeka lapho sisuswa endaweni yokugcina initrogen ewuketshezi, kudaleke ingozi kuyo yomibili imfucumfucu endizayo kanye nokuchayeka kokuqukethwe (1, 4).

Ngenxa yalezi zizathu, ukugcinwa kwezinga lokushisa eliphansi kakhulu kuvamise ukuba yi-nitrogen yesigaba somhwamuko. Lapho amasampula kufanele agcinwe esigabeni se-liquid, kufanele kusetshenziswe ishubhu ekhethekile ye-cryoflex.

Ububi besigaba somhwamuko ukuthi izinga lokushisa elime mpo lingenzeka okuholela ekushintshashintsheni kwezinga lokushisa phakathi kuka -135℃ kanye -190℃. Lokhu kudinga ukuqapha ngokucophelela nangenkuthalo amazinga e-nitrogen ewuketshezi kanye nokwehluka kwezinga lokushisa (5).

Abakhiqizi abaningi batusa ukuthi ama-cryovials afanele ukugcinwa phansi aze afike ku -135℃ noma asetshenziswe esigabeni somhwamuko kuphela.

Ukuncibilikisa Amaseli Akho E-Cryopreserved

Inqubo yokuncibilikisa icindezela isiko eliqandisiwe, futhi ukuphatha ngendlela efanele kanye nendlela edingekayo ukuze kuqinisekiswe ukusebenza kahle, ukululama, nokusebenza kwamaseli. Amaphrothokholi okuncibilika ngqo azoncika ezinhlotsheni ezithile zamaseli. Kodwa-ke, ukuncibilika ngokushesha kubhekwa njengokujwayelekile:

• Yehlisa noma imuphi umthelela ekululameni kweselula
• Siza ukunciphisa isikhathi sokuchayeka kuzincibilikisi ezikhona kumidiya eqandayo
• Nciphisa noma imuphi umonakalo ngokwenza kabusha amakristalu eqhwa

Amabhavu okugeza amanzi, okugeza ngobuhlalu, noma izinsimbi ezikhethekile ezizenzakalelayo zivame ukusetshenziselwa ukuncibilikisa amasampula.

Ezikhathini eziningi ulayini weseli o-1 uyancibilika ngesikhathi imizuzu engu-1-2, ngokushwiba kancane emanzini okugeza angu-37℃ kuze kube yilapho kusele iqhwa elincane ku-vial ngaphambi kokuba zigezwe endaweni yokukhula efudunyeziwe.

Kwamanye amangqamuzana anjengemibungu encelisayo, ukufudumala kancane kubalulekile ukuze aphile.

Amaseli manje alungele isiko lamaseli, ukuhlukaniswa kwamaseli, noma esimweni se-hematopoietic stem cell - izifundo ezisebenza kahle ukuze kuqinisekiswe ubuqotho bamaseli anikelayo ngaphambi kokwelashwa kwe-myeloablative.

Kuwumkhuba ovamile ukuthatha ama-aliquots amancane esampula eliwashwe ngaphambili elisetshenziselwa ukwenza isibalo samaseli ukuze kunqunywe ukugxiliswa kwamaseli ukuze kuhlanganiswe kusiko. Ungabe usuhlola imiphumela yezinqubo zokuhlukaniswa kwamaseli futhi unqume ukusebenza kweseli.

Imikhuba Engcono Kakhulu Yokugcina I-Cryovials

I-cryopreservation eyimpumelelo yamasampuli agcinwe kuma-cryovials kuncike ezintweni eziningi kuphrothokholi okuhlanganisa ukugcinwa okufanele nokugcinwa kwamarekhodi.

• Hlukanisa amaseli phakathi kwezindawo zokugcina– Uma imiqulu ivuma, hlukanisa amaseli phakathi kwamabhodlela futhi uwagcine ezindaweni ezihlukene ukuze unciphise ingozi yokulahleka kwesampula ngenxa yokwehluleka kwemishini.
• Vimbela ukungcoliswa okuphambene- Khetha ukusebenzisa kanye izitsha ze-cryogenic eziyinyumba noma i-autoclave ngaphambi kokusetshenziswa okulandelayo
• Sebenzisa izitsha ezinosayizi ofanele kumaseli akho- izitsha zifika ngohlu lwamavolumu phakathi kuka-1 no-5mls. Gwema ukugcwalisa izitsha ukuze unciphise ingozi yokuqhekeka.
• Khetha izitsha ze-cryogenic zangaphakathi noma zangaphandle- Izitsha ezifakwe ngaphakathi zituswa amanye amanyuvesi ngezinyathelo zokuphepha - zingavimbela futhi ukungcola ngesikhathi zigcwaliswa noma lapho zigcinwe ku-nitrogen ewuketshezi.
• Vimbela Ukuvuza- Sebenzisa ama-seal afakwe kabili abunjwe abe isikulufu-cap noma ama-O-rings ukuvimbela ukuvuza nokungcoliswa.
• Sebenzisa amabhakhodi e-2D namalebula amabhodlela- ukuqinisekisa ukulandeleka, izitsha ezinezindawo zokubhala ezinkulu zenza ukuthi ibhodlela ngalinye libhalwe ngokwanele. Amabhakhodi e-2D angasiza ngokuphathwa kwesitoreji nokugcinwa kwamarekhodi. Amakepisi anekhodi yombala awusizo ekuhlonzweni okulula.
• Ukugcinwa okwanele kwesitoreji- Ukuqinisekisa ukuthi amaseli awalahleki, imikhumbi yokugcina kufanele ihlale iqapha izinga lokushisa kanye namazinga e-nitrogen ewuketshezi. Ama-alamu kufanele afakwe ukuze azise abasebenzisi ngamaphutha.

Izinyathelo Zokuphepha

I-nitrogen ewuketshezi isiphenduke umkhuba ovamile ocwaningweni lwanamuhla kodwa inengozi yokulimala kabi uma isetshenziswa ngokungalungile.

I-personal protective equipment (PPE) kufanele igqokwe ukuze kuncishiswe ingozi yesithwathwa, ukusha nezinye izigameko ezimbi lapho uphatha i-nitrogen ewuketshezi. Gqoka

• Amagilavu ​​e-Cryogenic
• Ijazi laselabhorethri
• Isivikelo sobuso esiqinile esingazweli naso esimboze intamo
• Izicathulo ezivaliwe
• I-apron yepulasitiki ye-splashproof

Iziqandisi ze-nitrogen eziwuketshezi kufanele zibekwe ezindaweni ezinomoya ohlanzekile ukuze kuncishiswe ingozi yokuphelelwa umoya - i-nitrogen ephunyukile iyahwamuka futhi isuse umoya-mpilo osemkhathini. Izitolo ezinkulu zevolumu kufanele zibe nezinhlelo eziphansi ze-alamu ye-oxygen.

Ukusebenza ngababili lapho uphatha i-nitrogen ewuketshezi kuhle futhi ukusetshenziswa kwayo ngaphandle kwamahora okusebenza ajwayelekile kufanele kwenqatshelwe.

I-Cryovials yokusekela ukuhamba kwakho komsebenzi

Inkampani ye-Suzhou Ace Biomedical inikeza ukukhethwa okubanzi kwemikhiqizo ehlangabezana nezidingo zakho ze-cryopreservation zezinhlobo ezahlukene zamaseli. Iphothifoliyo ihlanganisa uhla lwamashubhu kanye nohlu lwama-cryovial oyinyumba.

Ama-cryovials ethu afaka:

• I-Lab Screw Cap 0.5mL 1.5mL 2.0mL Cryovial Cryogenic Vials Conical Bottom Cryotube with Gasket

  ● 0.5ml,1.5ml,2.0ml ukucaciswa, ngesiketi noma ngaphandle kwesiketi
  ● Idizayini eqinile noma ezimele, eyinyumba noma engeyona inyumba yomibili iyatholakala
  ● Amashubhu e-screw cap enziwe nge-polypropylene yebanga lezokwelapha
  ● Izitsha ze-PP Cryotube zingafrizwa ngokuphindaphindiwe futhi zincibilikiswe
  ●Idizayini yekhephu yangaphandle inganciphisa amathuba okungcola phakathi nesampula yokwelashwa.
  ● Isikulufa sekhephu yamashubhu e-cryogenic Imicu yesikulufu esijwayelekile ukuze isetshenziswe
  ● Amashubhu alingana nama-rotor avamile
  ● Amashubhu e-Cryogenic tube o-ring alingana ne-intshi engu-1 intshi no-2-inch, 48well, 81well, 96well kanye namabhokisi efriji angu-100well
  ● Ikwazi ukuzikhanyela ku-121°C futhi iqhwa ku- -86°C

  INGXENYE NO	IZINTO	UMQULU	I-CAPUMBALA	PCS/ISIKHWAMA	Izikhwama/Icala
  I-ACT05-BL-N	PP	0.5ML	Black,Yellow,Blue,Red,Purple,White	500	10
  I-ACT15-BL-N	PP	1.5ML	Black,Yellow,Blue,Red,Purple,White	500	10
  I-ACT15-BL-NW	PP	1.5ML	Black,Yellow,Blue,Red,Purple,White	500	10
  I-ACT20-BL-N	PP	2.0ML	Black,Yellow,Blue,Red,Purple,White	500	10