Li-Cryovialshangata li sebelisoa bakeng sa polokelo ea cryogenic ea mela ea lisele le lisebelisoa tse ling tsa bohlokoa tsa baeloji, li-dewars tse tlatsitsoeng ke naetrojene e mokelikeli.

Ho na le methati e 'maloa e amehang ho bolokeheng ka katleho liseleng tsa naetrojene e metsi. Le ha molao-motheo o le ho hoama butle, mokhoa o nepahetseng o sebelisitsoeng o ipapisitse le mofuta oa sele le cryoprotectant e sebelisitsoeng. Ho na le lintlha tse 'maloa tsa polokeho le mekhoa e metle eo u lokelang ho e ela hloko ha u boloka lisele ka mocheso o tlase joalo.

Poso ena e ikemiselitse ho fana ka kakaretso ea hore na li-cryovial li bolokoa joang ka har'a naetrojene e metsi.

Cryovials ke eng?

Li-cryovial ke linkho tse nyane, tse koaletsoeng tse etselitsoeng ho boloka lisampole tsa mokelikeli mochesong o tlase haholo. Li etsa bonnete ba hore lisele tse bolokiloeng ka cryoprotectant ha li kopane ka ho toba le metsi a naetrojene, ho fokotsa kotsi ea ho robeha ha lisele tsa cellular ha li ntse li rua molemo ho tsoa ho pholiso e feteletseng ea naetrojene ea metsi.

Hangata li-vial li fumaneha ka mefuta e mengata le meralo - e ka ba ka hare kapa ka ntle ho khoele ka likoti tse bataletseng kapa tse chitja. Liforomo tse sterile le tse sa hloekang le tsona lia fumaneha.

Ea SebelisangLi-Cyrovialsho Boloka Lisele ka Naetrojene e Metsi

Mefuta e mengata ea lilaboratori tsa NHS le tsa poraefete, hammoho le litsi tsa lipatlisiso tse sebetsanang le libanka tsa mali, epithelial cell biology, immunology le stem cell biology li sebelisa li-cryovials ho boloka lisele.

Lisele tse bolokiloeng ka tsela ena li kenyelletsa Lisele tsa B le T, Lisele tsa CHO, Lisele tsa Hematopoietic Stem le Progenitor Cells, Hybridomas, Intestinal Cells, Macrophages, Mesenchymal Stem le Progenitor Cells, Monocytes, Myeloma, NK Cells le Pluripotent Stem Cells.

Kakaretso ea Mokhoa oa ho boloka Cryovials ka Liquid Nitrogen

Cryopreservation ke ts'ebetso e bolokang lisele le lintho tse ling tsa tlhaho ka ho li pholisa ho mocheso o tlase haholo. Lisele li ka bolokoa ka har'a naetrojene e metsi ka lilemo ntle le tahlehelo ea matla a lisele. Ena ke kakaretso ea mekhoa e sebelisoang.

Ho Lokisetsa Lisele

Mokhoa o nepahetseng oa ho lokisa lisampole o tla fapana ho latela mofuta oa sele, empa ka kakaretso, lisele li bokelloa ebe li kenngoa bohareng ho hlahisa pellet e nang le lisele tse ngata. Pellet ena e khutlisetsoa ho supernatant e kopantsoeng le cryoprotectant kapa cryopreservation medium.

Cryopreservation Medium

Mokhoa ona o sebelisetsoa ho boloka lisele tse maemong a mocheso o tlase oo li tla ba tlas'a tsona ka ho thibela ho thehoa ha likristale tsa intracellular le extracellular, kahoo lisele li shoa. Karolo ea bona ke ho fana ka tikoloho e sireletsehileng, e sireletsang liseleng le liseleng nakong ea leqhoa, ho boloka le ho qhibiliha.

Mokelikeli o kang fresh frozen plasma (FFP), tharollo ea plasmalyte e nang le heparinised kapa serum-free, lisebelisoa tse se nang karolo ea liphoofolo li kopantsoe le li-cryoprotectants tse kang dimethyl sulphoxide (DMSO) kapa glycerol.

The re-liquified sample pellet is aliquoted into polypropylene cryovials such asKhamphani ea Suzhou Ace Biomedical Cryogenic Storage Vials.

Ke habohlokoa hore u se ke ua tlatsa li-cryovials kaha sena se tla eketsa kotsi ea ho phunyeha le ho lokolloa ha litaba (1).

Sekhahla sa Leqhoa le Laoloang

Ka kakaretso, sekhahla sa leqhoa se laoloang butle se sebelisoa bakeng sa ho boloka lisele tse atlehileng.

Ka mor'a hore lisampole li kenngoe ka har'a linkho tsa cryogenic, li behoa holim'a leqhoa le metsi kapa ka sehatsetsing sa 4 ℃ 'me mokhoa oa ho leqhoa o qala nakong ea metsotso e 5. Joaloka tataiso e akaretsang, lisele li pholile ka lebelo la -1 ho ea ho -3 ka motsotso (2). Sena se finyelloa ka ho sebelisa sehatsetsi se ka sebelisoang kapa ka ho beha linkho ka har'a sehatsetsi se laoloang sa -70 ° C ho ea -90 ° C.

Fetisetsa ho Liquid Nitrogen

Linkho tsa cryogenic tse hatselitsoeng li fetisetsoa ka tankeng ea metsi ea naetrojene ka nako e sa lekanyetsoang ha mocheso o ka tlase ho -135 ℃ o bolokoa.

Lithempereichara tsena tse tlase haholo li ka fumanoa ka ho qoelisoa ka har'a mokelikeli kapa mouoane oa naetrojene.

Phase ea Mokelikeli kapa Mouoane?

Ho boloka naetrojene ea mokelikeli e tsejoa ka ho boloka mocheso o batang ka mokhoa o tsitsitseng, empa hangata ha e khothalletsoe ka mabaka a latelang:

• Tlhokahalo ea bongata bo boholo (botebo) ba naetrojene e metsi e leng kotsi e ka bang teng. Ho chesa kapa ho felloa ke matla ka lebaka la sena ke kotsi ea sebele.
• Linyeoe tse ngolisitsoeng tsa tšoaetso ea tšoaetso ke likokoana-hloko tse kang aspergillus, hep B le ho ata ha kokoana-hloko ka mokhoa oa metsi oa nitrogen (2,3)
• Monyetla oa hore naetrojene e metsi e lutlele ka har'a linkho nakong ea ho qoelisoa. Ha e tlosoa polokelong 'me e futhumala ho ea mocheso oa kamore, naetrojene e hōla ka potlako. Ka lebaka leo, vial e ka 'na ea robeha ha e tlosoa polokelong ea naetrojene ea metsi, ea baka kotsi ho lithōle tse fofang le ho pepesehela lintho tse ka hare (1, 4).

Ka mabaka ana, ho boloka mocheso o tlase haholo hangata ho naetrojene ea mouoane. Ha lisampole li tlameha ho bolokoa molemong oa mokelikeli, ho lokela ho sebelisoa li-tubing tse khethehileng tsa cryoflex.

Taba e nyahamisang ea mohato oa mouoane ke hore mocheso o otlolohileng oa mocheso o ka hlaha o bakang ho fetoha ha mocheso pakeng tsa -135 ℃ le -190 ℃. Sena se hloka tlhokomeliso e hlokolosi le e hlokolosi ea maemo a mokelikeli a naetrojene le phapang ea mocheso (5).

Baetsi ba bangata ba khothaletsa hore li-cryovial li loketse ho bolokoa ho fihlela ho -135 ℃ kapa ho sebelisoa mokhahlelong oa mouoane feela.

Ho qhibiliha Lisele Tsa Hao Tse Cryopreserved

Ts'ebetso ea ho qhibiliha e sithabetsa moetlong o hatsetseng, 'me ho ts'oaroa le mokhoa o nepahetseng oa hlokahala ho netefatsa ts'ebetso e nepahetseng, ho hlaphoheloa le ho sebetsa ha lisele. Mekhoa e nepahetseng ea ho qhibiliha e tla itšetleha ka mefuta e itseng ea lisele. Leha ho le joalo, ho qhibiliha ka potlako ho nkoa e le ntho e tloaelehileng ho:

• Fokotsa tšusumetso leha e le efe ea ho hlaphoheloa ha lisele
• Thusa ho fokotsa nako ea ho pepeseha ho li-solutes tse teng mecheng ea litaba tse batang
• Fokotsa tšenyo leha e le efe ka ice recrystallization

Libate tsa metsi, libate tsa lifaha, kapa lisebelisoa tse ikhethileng tse sebelisoang hangata ho qhibilihisa lisampole.

Hangata mohala o le mong oa sele o qhibiliha ka nako bakeng sa metsotso e 1-2, ka ho phunya ka bonolo ka har'a bateng ea metsi ea 37℃ ho fihlela ho setse leqhoa le lenyenyane ka har'a vial pele e hlatsuoa ka mokhoa oa ho hōla o futhumetseng.

Bakeng sa lisele tse ling tse kang mahe a mammalian, ho futhumala butle ho bohlokoa bakeng sa ho phela.

Lisele li se li loketse moetlo oa lisele, ho aroloa ha lisele, kapa tabeng ea lisele tsa stem tsa haematopoietic - lithuto tsa ho sebetsa ho netefatsa botšepehi ba lisele tsa stem pele ho phekolo ea myeloablative.

Ke tloaelo e tloaelehileng ho nka li-aliquots tse nyane tsa sampole e hlatsoitsoeng e sebelisetsoang ho etsa palo ea lisele ho fumana hore na ke bokae ba lisele bakeng sa ho rala moetlong. Joale u ka lekola liphetho tsa mekhoa ea ho arola lisele le ho fumana hore na sele e sebetsa hantle hakae.

Mekhoa e metle ea ho boloka li-Cryovials

Cryopreservation e atlehileng ea lisampole tse bolokiloeng ka har'a li-cryovials e itšetlehile ka likarolo tse ngata tsa protocol ho kenyeletsa polokelo e nepahetseng le ho boloka tlaleho.

• Arola lisele pakeng tsa libaka tsa polokelo- Haeba meqolo e lumella, arola lisele pakeng tsa li-vials 'me u li boloke libakeng tse arohaneng ho fokotsa kotsi ea tahlehelo ea mohlala ka lebaka la ho hlōleha ha thepa.
• Thibela tšilafalo- Khetha bakeng sa tšebeliso e le 'ngoe ea linkho tsa cryogenic kapa li-autoclave pele u li sebelisa
• Sebelisa libotlolo tsa boholo bo lekaneng bakeng sa lisele tsa hau- Linkho li tla ka mefuta e mengata e pakeng tsa 1 le 5mls. Qoba ho tlatsa linkho ho fokotsa kotsi ea ho peperana.
• Khetha ka hare kapa ka ntle likhoele tsa cryogenic- Likotlolo tse kentsoeng ka har'a likhoele li khothaletsoa ke liunivesithi tse ling bakeng sa mehato ea polokeho - li ka thibela ts'oaetso nakong ea ho tlatsoa kapa ha li bolokoa ka har'a naetrojene e metsi.
• Thibela Leakage- Sebelisa litiiso tse kentsoeng ka makhetlo a mabeli tse entsoeng ka har'a screw-cap kapa O-rings ho thibela ho lutla le ho silafala.
• Sebelisa libarcode tsa 2D le libotlolo tsa mabitso- ho etsa bonnete ba ho ts'oara, linkho tse nang le libaka tse kholo tsa ho ngolla li thusa hore vial e 'ngoe le e 'ngoe e ngoloe ka mokhoa o lekaneng. Li-barcode tsa 2D li ka thusa ka taolo ea polokelo le ho boloka rekoto. Li-caps tsa mebala li bohlokoa bakeng sa ho tsebahatsa habonolo.
• Tlhokomelo e lekaneng ea polokelo- Ho etsa bonnete ba hore lisele ha li lahlehe, lijana tsa polokelo li lokela ho lula li lekola mocheso le maemo a naetrojene ea metsi. Lialamo li lokela ho kenngoa ho lemosa basebelisi ka liphoso.

Litlhokomelo tsa Tšireletso

Naetrojene e nang le metsi e se e tloaelehile liphuputsong tsa morao-rao empa e na le kotsi ea kotsi e tebileng haeba e sebelisoa ka mokhoa o fosahetseng.

Thepa e nepahetseng ea ho itšireletsa (PPE) e lokela ho apesoa ho fokotsa kotsi ea serame, ho cha le liketsahalo tse ling tse mpe ha ho sebelisoa naetrojene ea metsi. Apara

• Liatlana tsa cryogenic
• Seaparo sa laboratori
• Thebe e felletseng ea sefahleho e hananang le tšusumetso e koahelang molala
• Lieta tse koetsoeng
• Apron ea polasetiki ea Splashproof

Lihatsetsi tsa naetrojene tse metsi li lokela ho behoa libakeng tse nang le moea o motle ho fokotsa kotsi ea ho felloa ke moea - naetrojene e phonyohileng ea fetoha mouoane ebe e tlosa oksijene ea sepakapaka. Mabenkele a maholo a mangata a lokela ho ba le lisebelisoa tse fokolang tsa oksijene.

Ho sebetsa ka bobeli ha u tšoara naetrojene e nang le metsi ho loketse 'me tšebeliso ea eona ka ntle ho lihora tse tloaelehileng tsa ho sebetsa e lokela ho thibeloa.

Li-Cryovials ho Tšehetsa Ts'ebetso ea Hao ea Mosebetsi

Khamphani ea Suzhou Ace Biomedical e fana ka khetho e pharaletseng ea lihlahisoa tse fihlelang litlhoko tsa hau tsa cryopreservation bakeng sa mefuta e fapaneng ea lisele. Potefolio e kenyelletsa mefuta e mengata ea li-tubesas hammoho le mefuta e mengata ea li-cryovial tse nyopa.

Li-cryovials tsa rona ke tsena:

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

  ● 0.5ml, 1.5ml, 2.0ml litlhaloso, ka moleka kapa ntle le mose
  ● Moetso oa conical kapa oa ho ikemela, o hloekile kapa o sa hloeka ka bobeli li teng
  ● Li-tubes tsa Screw cap li entsoe ka polypropylene ea boemo ba bongaka
  ● Likotlolo tsa PP Cryotube li ka qhomisoa khafetsa le ho qhibiliha
  ● Sebopeho sa cap e ka ntle se ka fokotsa monyetla oa tšoaetso nakong ea phekolo ea mohlala.
  ● Screw cap cryogenic tubes Universal screw threads bakeng sa tšebeliso
  ● Li-tubes li lumellana le li-rotor tse tloaelehileng
  ● Cryogenic tube o-ring tubes e lekana le mabokose a sehatsetsi a 1-inch le 2-inch, 48well, 81well,96well le 100well.
  ● E khona ho totobala ho 121°C 'me e ka hatsetsa ho -86°C

  KAROLO NO	TŠEBELETSO	BOLUMO	CAPMOLAOLI	PCS/MOKOTLA	MKOTLA/NYENYANE
  ACT05-BL-N	PP	0.5ML	Botsho, Bosehla, Botala, Bofubedu, Pherese, Bosweu	500	10
  ACT15-BL-N	PP	1.5ML	Botsho, Bosehla, Botala, Bofubedu, Pherese, Bosweu	500	10
  ACT15-BL-NW	PP	1.5ML	Botsho, Bosehla, Botala, Bofubedu, Pherese, Bosweu	500	10
  ACT20-BL-N	PP	2.0ML	Botsho, Bosehla, Botala, Bofubedu, Pherese, Bosweu	500	10