KryovialHoʻohana maʻamau ʻia no ka mālama ʻana i ka cryogenic o nā laina kelepona a me nā mea koʻikoʻi koʻikoʻi koʻikoʻi, i loko o nā dewar i hoʻopiha ʻia me ka wai nitrogen.

Nui nā ʻanuʻu i ka mālama ʻana i nā cell i loko o ka wai nitrogen. ʻOiai ʻo ka loina kumu he lohi lohi, ʻo ka ʻenehana pololei i hoʻohana ʻia ma muli o ke ʻano cell a me ka cryoprotectant i hoʻohana ʻia. Nui nā noʻonoʻo palekana a me nā hana maikaʻi loa e noʻonoʻo i ka wā e mālama ai i nā cell i kēlā me kēia haʻahaʻa haʻahaʻa.

Ke manaʻo nei kēia pou e hāʻawi i kahi ʻike o ka mālama ʻana i nā cryovial i loko o ka wai nitrogen.

He aha nā Cryovial

He liʻiliʻi nā kīʻaha Cryovial i hoʻopaʻa ʻia i hoʻolālā ʻia no ka mālama ʻana i nā mea hoʻohālike wai ma nā wela haʻahaʻa loa. Hoʻopaʻa lākou ʻaʻole pili pono nā cell i mālama ʻia i loko o ka cryoprotectant me ka nitrogen wai, e hōʻemi ana i ka pilikia o ka haki ʻana o nā kelepona ʻoiai e pōmaikaʻi ana i ka hopena hoʻoluʻu loa o ka nitrogen wai.

Loaʻa pinepine ʻia nā hue i loko o nā ʻano nui a me nā hoʻolālā - hiki ke hoʻopaʻa ʻia i loko a i waho paha me nā lalo palahalaha a poepoe paha. Loaʻa ʻia nā ʻano hoʻopono a me nā ʻano ʻole.

Na wai e hoohanaʻO Cyroviale mālama i nā pūnaewele i loko o ka Nitrogen Liquid

ʻO kahi ʻano o ka NHS a me nā hale hana pilikino, a me nā keʻena noiʻi kūikawā i ka panakō koko kaula, epithelial cell biology, immunology a me stem cell biology e hoʻohana i nā cryovial i cryopreserve cell.

ʻO nā pūnaewele mālamaʻia ma kēiaʻano,ʻo ia nā pūnaewele B a me T, CHO Cells, Hematopoietic Stem and Progenitor Cells, Hybridomas, Intestinal Cells, Macrophages, Mesenchymal Stem and Progenitor Cells, Monocytes, Myeloma, NK Cells and Pluripotent Stem Cells.

ʻO ka nānā ʻana o ka mālama ʻana i nā Cryovial i loko o ka Nitrogen Liquid

ʻO ka cryopreservation kahi kaʻina e mālama ai i nā cell a me nā mea olaola ʻē aʻe ma o ka hoʻomaha ʻana iā lākou i nā wela haʻahaʻa loa. Hiki ke mālama ʻia nā kelepona i loko o ka nitrogen wai no nā makahiki me ka ʻole o ka nalowale o ke ola o ke kelepona. He papa kuhikuhi kēia o nā kaʻina hana i hana ʻia.

Hoʻomākaukau Cell

E ʻokoʻa ke ala pololei no ka hoʻomākaukau ʻana i nā laʻana ma muli o ke ʻano cell, akā ma ka laulā, ʻohi ʻia nā cell a centrifuged e hoʻomohala i kahi pellet waiwai nui. Hoʻopili hou ʻia kēia pellet i loko o ka supernatant i hui pū ʻia me ka cryoprotectant a i ʻole kahi ʻano cryopreservation.

Cryopreservation Medium

Hoʻohana ʻia kēia ʻano no ka mālama ʻana i nā cell i nā wahi haʻahaʻa haʻahaʻa e hoʻokau ʻia ai lākou ma ke kāohi ʻana i ke kūkulu ʻia ʻana o nā kristal intra a me extracellular a no laila ka make cell. ʻO kā lākou kuleana, ʻo ia ka hāʻawi ʻana i kahi wahi palekana, palekana no nā cell a me nā ʻiʻo i ka wā o ka hoʻoheheʻe ʻana, ka mālama ʻana, a me ka hoʻoheheʻe ʻana.

Hoʻohui ʻia kahi ʻano e like me ka plasma hau hau hou (FFP), heparinised plasmalyte solution a i ʻole serum-free, nā ʻāpana holoholona ʻole i hui pū ʻia me nā cryoprotectants e like me dimethyl sulphoxide (DMSO) a i ʻole glycerol.

Hoʻokaʻawale ʻia ka pellet hāpana hou i loko o nā cryovial polypropylene e like meʻO Suzhou Ace Biomedical hui Cryogenic Storage Vials.

He mea nui ʻaʻole e hoʻopiha piha i nā cryovial no ka mea e hoʻonui kēia i ka pilikia o ka haki ʻana a me ka hoʻokuʻu ʻana o nā mea i loko (1).

Ka Hoʻopaʻa Paʻa Paʻa

Ma keʻano laulā, hoʻohana ʻia kahi helu hoʻokuʻu lohi no ka cryopreservation kūleʻa o nā cell.

Ma hope o ka hoʻokaʻawale ʻia ʻana o nā laʻana i loko o nā hue cryogenic, kau ʻia lākou ma luna o ka hau mākū a i ʻole i loko o kahi pahu hau 4 ℃ a hoʻomaka ke kaʻina hana manuahi i loko o 5 mau minuke. Ma ke ʻano he alakaʻi maʻamau, hoʻomaʻalili ʻia nā cell ma ka wikiwiki o -1 a -3 i kēlā me kēia minuke (2). Hoʻokō ʻia kēia me ka hoʻohana ʻana i kahi mea hoʻomaʻamaʻa programmable a i ʻole ma ka waiho ʻana i nā hue i loko o kahi pahu insulated i hoʻokomo ʻia i loko o ka pahu hau hoʻokele -70°C a i -90°C.

Hoʻololi i ka Liquid Nitrogen

Hoʻololi ʻia nā hue cryogenic maloʻo i loko o kahi pahu wai nitrogen wai no ka manawa pau ʻole i mālama ʻia ka mahana ma lalo o -135 ℃.

Hiki ke loaʻa kēia mau wela haʻahaʻa loa ma o ka hoʻokomo ʻia ʻana i loko o ka wai a i ʻole ka vapor phase nitrogen.

ʻO ka wai a i ʻole ka ʻauwa?

ʻIke ʻia ka mālama ʻana i loko o ka wai hau hau no ka mālama ʻana i ke anuanu me ka paʻa paʻa, akā ʻaʻole ʻōlelo pinepine ʻia no kēia mau kumu:

• ʻO ka pono o nā puke nui (hohonu) o ka waiʻona wai he mea pōʻino. ʻO ke ahi a i ʻole ka asphyxiation ma muli o kēia he pilikia maoli.
• Hoʻopaʻa ʻia nā hihia o ka hoʻohaumia ʻana e nā mea maʻi maʻi e like me ka aspergillus, hep B a me ka hoʻolaha ʻana o ka maʻi viral ma o ka wai nitrogen (2,3)
• ʻO ka hiki ke hoʻoheheʻe ʻia ka nitrogen wai i loko o nā hue i ka wā o ka hoʻoinu ʻana. Ke hoʻoneʻe ʻia mai ka waiho ʻana a hoʻomaʻamaʻa ʻia i ka lumi wela, hoʻonui wikiwiki ka nitrogen. No laila, nahaha paha ka hue i ka wā e wehe ʻia ai mai kahi waiho ʻana o ka nitrogen wai, e hana ana i kahi pōʻino mai nā ʻōpala lele a me ka ʻike ʻana i nā mea i loko (1, 4).

No kēia mau kumu, ʻoi aku ka maʻamau o ka mālama ʻana i ka wela haʻahaʻa loa i loko o ka vapor phase nitrogen. Ke mālama ʻia nā laʻana i loko o ka pae wai, pono e hoʻohana ʻia nā paipu cryoflex kūikawā.

ʻO ka ʻaoʻao haʻahaʻa i ka pae mahu, ʻo ia ka hiki ʻana o kahi ʻoniʻoni wela kū i ka hopena i ka loli ʻana o ka mahana ma waena o -135 ℃ a me -190 ℃. Pono kēia i ka nānā pono ʻana a me ka hoʻoikaika ʻana i nā pae wai a me nā ʻano like ʻole o ka mahana (5).

Manaʻo ka nui o nā mea hana i kūpono nā cryovial no ka mālama ʻana a hiki i -135 ℃ a i ʻole no ka hoʻohana ʻana i ka pae mahu wale nō.

Ka hoʻoheheʻe ʻana i kāu mau pūnaewele Cryopreserved

He mea koʻikoʻi ke kaʻina hana hoʻoheheʻe no ka moʻomeheu hau, a pono ka lawelawe ʻana a me ka ʻenehana no ka hōʻoia ʻana i ka hiki ke ola, hoʻihoʻi hou, a me ka hana o nā cell. E hilinaʻi nā protocols thawing pololei i nā ʻano kelepona kikoʻī. Eia naʻe, ua manaʻo ʻia ka hoʻoheheʻe wikiwiki ʻana i ka maʻamau i:

• E ho'ēmi i kekahi hopena i ka hoʻihoʻi kelepona
• E kōkua i ka hoʻemi ʻana i ka manawa hoʻolaha i nā solute i loaʻa i loko o ka media maloʻo
• E hōʻemi i ka pōʻino ma muli o ka hana hou ʻana o ka hau

Hoʻohana pinepine ʻia nā ʻauʻau wai, nā ʻauʻau ʻauʻau, a i ʻole nā ​​​​mea hana ʻenehana kūikawā no ka hoʻoheheʻe ʻana i nā laʻana.

ʻO ka maʻamau, hoʻoheheʻe ʻia ka laina kelepona 1 i ka manawa no 1-2 mau minuke, ma ka wili mālie ʻana i loko o kahi ʻauʻau wai 37 ℃ a hiki i ka waiho ʻana o kahi hau liʻiliʻi i loko o ka hue ma mua o ka holoi ʻia ʻana i loko o kahi ʻano ulu i hoʻomaʻamaʻa ʻia.

No kekahi mau pūnaewele e like me nā embryos mammalian, pono ka hoʻomehana lohi no ko lākou ola.

Ua mākaukau nā pūnaewele no ka moʻomeheu cell, ka hoʻokaʻawale ʻana i ke kelepona, a i ʻole ke ʻano o ka haematopoietic stem cell - nā haʻawina ola e hōʻoiaʻiʻo i ka kūpaʻa o nā mea hāʻawi pū ma mua o ka myeloablative therapy.

He hana maʻamau ka lawe ʻana i nā liʻiliʻi liʻiliʻi o ka laʻana i holoi ʻia i hoʻohana ʻia e hana i ka helu kelepona no ka hoʻoholo ʻana i nā manaʻo o nā cell no ka hoʻopili ʻana i ka moʻomeheu. A laila hiki iā ʻoe ke loiloi i nā hopena o nā kaʻina hana hoʻokaʻawale cell a hoʻoholo i ka ola ola.

Nā hana maikaʻi loa no ka mālama ʻana i nā Cryovial

ʻO ka holomua cryopreservation o nā laʻana i mālama ʻia i loko o nā cryovial e pili ana i nā mea he nui o ka protocol me ka mālama pono a me ka mālama ʻana i nā moʻolelo.

• E hoʻokaʻawale i nā pūnaewele ma waena o nā wahi mālama- Inā ʻae ka nui, e hoʻokaʻawale i nā cell ma waena o nā hue a mālama iā lākou i nā wahi ʻokoʻa e hōʻemi i ka hopena o ka nalowale o ka laʻana ma muli o nā hemahema o nā lako.
• Kāohi i ka hoʻohaumia ʻana- E koho i nā hue cryogenic sterile hoʻohana hoʻokahi a i ʻole autoclave ma mua o ka hoʻohana ʻana
• E hoʻohana i nā hue wai kūpono no kāu mau keena- hele mai nā hue i loko o ka nui o nā puke ma waena o 1 a me 5mls. E hōʻalo i ka hoʻopiha piha ʻana i nā hue i mea e hōʻemi ai i ka pōʻalo.
• E koho i nā hue cryogenic i loko a i ʻole waho- Manaʻo ʻia nā hue i hoʻopaʻa ʻia i loko e kekahi mau kulanui no nā hana palekana - hiki iā lākou ke pale i ka ʻino i ka wā o ka hoʻopiha ʻana a i ʻole ke mālama ʻia i loko o ka wai nitrogen.
• Kāohi i ka Leakage- E hoʻohana i nā hōʻailona bi-injected i hoʻoheheʻe ʻia i loko o ka pōʻai screw a i ʻole O-rings e pale ai i ka lī a me ka haumia.
• E hoʻohana i nā barcode 2D a me nā hue lepili- no ka hōʻoia ʻana i ka traceability, nā hue me nā wahi kākau nui e hiki ai i kēlā me kēia hue ke hōʻailona kūpono. Hiki i nā barcode 2D ke kōkua i ka mālama ʻana a me ka mālama moʻolelo. Pono nā pāpale kala kala no ka ʻike maʻalahi.
• ʻO ka mālama mālama pono- No ka hōʻoia ʻana ʻaʻole nalowale nā ​​​​pūnaewele, pono e nānā mau nā moku mālama i ka mahana a me nā pae wai nitrogen. Pono e hoʻopaʻa ʻia nā ʻōlima e hoʻomaopopo i nā mea hoʻohana i nā hewa.

Palekana Palekana

Ua lilo ka waiona wai i mea maʻamau i ka noiʻi ʻana i kēia wā akā hiki ke loaʻa i ka pōʻino koʻikoʻi inā hoʻohana hewa ʻia.

Pono e hoʻohana pono i nā lako pale pilikino (PPE) e hōʻemi i ka pilikia o ka hau hau, ke kuni a me nā mea ʻino ʻē aʻe i ka wā e lawelawe ai i ka nitrogen wai. ʻaʻahu

• Nā mīkina lima cryogenic
• ʻAʻahu Laboratory
• ʻO ka pale pale maka piha i ka hopena e uhi pū ana i ka ʻāʻī
• Nā kāmaʻa pani
• ʻAiʻi paʻi paʻipaʻi

Pono e hoʻokomo ʻia nā pahu hau hau hau i loko o nā wahi i hoʻoluʻu maikaʻi ʻia e hōʻemi i ka pilikia o ka asphyxiation - hoʻokuʻu ʻia ka nitrogen a hoʻoneʻe i ka oxygen atmospheric. Pono nā hale kūʻai leo nui e loaʻa nā ʻōnaehana alarm oxygen haʻahaʻa.

He mea maikaʻi ka hana ʻelua i ka wā e lawelawe ai i ka nitrogen wai a pāpā ʻia ka hoʻohana ʻana ma waho o nā hola hana maʻamau.

Nā Cryovial e kākoʻo i kāu Kaʻina Hana

Hāʻawi ka hui ʻo Suzhou Ace Biomedical i kahi koho nui o nā huahana e hoʻokō i kāu pono cryopreservation no nā ʻano cell like ʻole. Aia i loko o ka waihona ka nui o nā tubesas a me nā ʻano cryovial sterile.

ʻO kā mākou cryovial:

• Paʻi Screw Lab 0.5mL 1.5mL 2.0mL Cryovial Cryogenic Vials Conical lalo Cryotube me Gasket

  ● 0.5ml,1.5ml,2.0ml kiko'ī, me ke kapa a me ke kapa ole
  ● Hoʻolālā conical a kū ponoʻī paha, sterile a i ʻole sterile nā ​​mea ʻelua i loaʻa
  ● Ua hana ʻia nā paipu pōleʻa me ka polypropylene lāʻau lapaʻau
  ● Hiki i ka PP Cryotube Vials ke hoʻomaloʻo pinepine ʻia a hoʻoheheʻe ʻia
  ● Hiki i ka hoʻolālā pāpale waho ke hōʻemi i ka hiki ke hoʻohaumia i ka wā o ka lāʻau lapaʻau.
  ● Poʻo uʻi cryogenic paipu Uila wili honua no ka hoʻohana ʻana
  ● Pono nā paipu i nā rotors maʻamau
  ● Paipu Cryogenic o-ring paipu i kūpono i nā pahu pahu hau 1-inihi a me 2-inihi, 48well, 81well，96well a me 100well.
  ● Hiki i ka autoclavable a hiki i 121°C a hiki i ka -86°C

  MAHELE NO	KA MEA	NUI	KAPAKULA	PCS/ʻEKE	NA EKE/PAE
  HANA05-BL-N	PP	0.5ML	ʻeleʻele, melemele, uliuli, ʻulaʻula, poni, keʻokeʻo	500	10
  HANA15-BL-N	PP	1.5ML	ʻeleʻele, melemele, uliuli, ʻulaʻula, poni, keʻokeʻo	500	10
  HANA15-BL-NW	PP	1.5ML	ʻeleʻele, melemele, uliuli, ʻulaʻula, poni, keʻokeʻo	500	10
  HANA20-BL-N	PP	2.0ML	ʻeleʻele, melemele, uliuli, ʻulaʻula, poni, keʻokeʻo	500	10