Pehea e hoʻohana ai i ka paipu cryopreservation me ka ʻepekema a me ka pololei
He ʻepekema ka hoʻohana ʻana i ka paipu cryopreservation, ʻaʻole he trilogy maʻalahi e like me ka wehe ʻana i ka pahu wai nitrogen wai, hoʻokomo i loko o ka pahu cryopreservation a pani i ka pahu hau hau.Hiki i ka ʻepekema a me ka hoʻohana pololei ʻana i nā paipu cryopreservation ke pale i ka nalowale o nā laʻana a pale i ka palekana o nā mea hōʻike.
He ʻepekema ka hoʻohana ʻana i ka paipu cryopreservation, ʻaʻole he trilogy maʻalahi e like me ka wehe ʻana i ka pahu wai nitrogen wai, hoʻokomo i loko o ka pahu cryopreservation a pani i ka pahu hau hau.Hiki i ka ʻepekema a me ka hoʻohana pololei ʻana i nā paipu cryopreservation ke pale i ka nalowale o nā laʻana a pale i ka palekana o nā mea hōʻike.
Paipu hau: ʻanuʻu hau
Holoi i na keena me ka wai PBS preheated, omo i ka wai hooluu, a uhi i na keena me ka solution i loko o trypsin a me EDTA (ua lawa ka wai wai lahilahi, a pono e hooholo i ka manao o trypsin a me EDTA e like me ka laina cell).
E hoʻomoʻa i nā cell ma 37 ℃ no 3-5 mau minuke.
Ma hope o ka haʻalele ʻana o nā cell mai lalo mai, hoʻopau ʻia ka incubation, hoʻohui ʻia ka medium i loko o ka serum, a hoʻokuʻu mālie ʻia nā cell me kahi pipette.
E Centrifuge i ka hoʻokuʻu ʻana o ke kelepona (500 xg, 5 min) a me ka resuspension me ka mea e loaʻa ai ka serum.
helu kelepona.
E centrifuge i ka hoʻokuʻu ʻana o ke kelepona (500 xg, 5 mau minuke), e hoʻoneʻe i ka supernatant, a e hoʻihoʻi hou i nā cell me ka medium i loaʻa ka serum o ka nui kūpono.
E hoʻohui i nā cell a me ka cryopreservation solution (60% medium, 20% fetal bovine serum, 20% DMSO) i 1:1 volume ratio, a laila e hoʻoili iā lākou i ka Cryo STM cryopreservation tube.He 1-5 × 106 mau ʻāpana/ml ka nui o nā pūnae hau.
Manaʻo ʻia ʻo Cryo e loaʻa ana i nā cell STM cryopreservation tube e hoʻomaloʻo i ka wikiwiki o −1 K/min, a hiki ke waiho ʻia ka paipu cryopreservation i loko o kahi pahu i loaʻa ka isopropanol ma − 70 ℃.Inā mālama ʻo Cryo STM cryopreservation tube i nā laʻana ʻē aʻe, hiki ke waiho pololei ʻia ma − 20 ℃, − 70 ℃ a i ʻole ka pae kinoea o ka nitrogen wai.No ka hoʻopaʻa ʻana i ka paʻa like ʻana o ka hāpana, 4 mL a me 5 mL Cryo Pono e hoʻokomo ʻia ka paipu cryopreservation sTM i loko o ka friji ma -20 ℃ no ka pō, a laila e hoʻoneʻe ʻia i ka manawa kinoea o - 70 ℃ a i ʻole ka wai hau.
A laila e hoʻololi i ka paipu cryopreservation Cryo.sTM i ka pahu hau hau.I mea e pale aku ai i ka pollution (e like me ka mycoplasma) a me ka noʻonoʻo ʻana i ka palekana, e ʻoluʻolu e kau i ka paipu cryopreservation Cryo.sTM i loko o ka māhele kinoea o ka naikokene wai, ʻaʻole i ka pae wai.
Pehea e hoʻohana ai i ka paipu cryopreservation me ka ʻepekema a me ka pololei?Hoʻokumu ʻia kā mākou hui i nā poʻe loea me ka ʻoihana ʻoihana a me ka ʻike mākeke waiwai e hoʻolako i nā huahana no ke kahua noiʻi ʻepekema ola a me nā lawelawe no nā mea noiʻi ʻepekema.ʻAʻole hiki iā ia ke hoʻokō wale i nā koi kūikawā o nā mea kūʻai aku R&D ma nā ʻano huahana a me ka hoʻopili ʻana, akā hoʻokō pū i nā pono piha o nā ʻoihana hana i nā pae āpau mai ka liʻiliʻi liʻiliʻi, ka nui waena a hiki i ka hana nui.
Ka manawa hoʻouna: Nov-25-2022