I-AMMS®NK Cell Culture Kit 3.0

I-Natural Killer (NK) yokwelapha ngamaseli isiphenduke inkomba yocwaningo olugxile kuyo ekuhlolweni kokuzivikela komzimba, okubonisa amandla amangalisayo ku-oncology. Amaseli e-NK angama-lymphocyte azalwa nawo anomsebenzi obanzi we-cytotoxic, akwazi ukukhomba amaseli wesimila, amaseli atheleleke ngegciwane, kanye nezinye izifo. Ihlukile emithi yokwelapha evamile ye-T cell, amaseli e-NK asebenzisa imiphumela esheshayo ye-cytotoxic ngaphandle kokuqala kwe-antigen, okuvumela isenzo sokwelapha esisheshayo esizimele ngaphandle kokucaciswa kwe-tumor antigen.

Umfanekiso 1. Izindlela zokulwa nesimila zamaseli e-NK
(Umthombo wesithombe: Acta Pharmaceutica Sinica)

Amaseli e-NK angatholakala emisuka eyahlukene, okuhlanganisa igazi le-peripheral, igazi lenkaba (UCB), amaseli e-pluripotent stem cell (iPSCs), kanye nomugqa weseli we-NK92. Ucwaningo lugqamisa ukuthi amaseli e-NK atholakala ku-UCB kanye nokhokho babonisa inala ephezulu kakhulu phakathi kwemithombo etholakalayo. I-naïve phenotype yamaseli e-NK asuselwa ku-UCB anciphisa ubungozi be-Graft-versus-Host Disease (GVHD), kuyilapho ukutholakala kwawo okubanzi kanye nesikhundla esiphakeme sokwandisa umthamo we-UCB njengomthombo wamasu wokukhiqiza amaseli e-NK. Ngokuphawulekayo, isilingo sokuqala somtholampilo se-CAR-NK sasebenzisa amaseli e-NK asuselwa ku-UCB. Ukugqama okukhulayo kwamaseli e-UCB-NK ocwaningweni lwe-immunotherapy kugcizelela inselele ebalulekile yokuzuza ukwanda kwamaseli e-NK ahlanzekile, amsulwa kakhulu.ku-vitroukuhumusha komtholampilo.

I-AMMS®NK Cell Culture Kit 3.0 yakhelwe ukusetshenziswa ex vivoukwenziwa kusebenze kanye nokwandiswa kwamaseli e-NK ahlanzekile kakhulu asuka kumaseli enkaba egazi amhlophe noma acwebezelayo e-mononuclear cell (CBMCs). Uhlelo lwesiko lwe-2-litre luzuza isivuno samaseli ayizigidigidi ezi-4-6 ngokuhlanzeka kwe-NK okwedlula u-90%.

.Izici Eziyinhloko.

• .I-Chemically defined Culture.
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• Ukuhambisana kwesampula: Ama-UCB CBMC amasha noma agcinwe e-cryopreserved.
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• Ukwakhiwa kwe-Xeno-free.
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• Ukunwetshwa kwesivuno esiphezulu: amaseli ayizigidigidi ezi-4-6 ngohlelo lwe-2L olune->90% NK ubumsulwa

Idatha Yokusebenza

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Umfanekiso1. Isamba sokunwetshwa kweseli nokusebenza kwamaseli e-NK asuselwa ku-UCB akhiwe nge-AMMS®NK Cell Culture Kit 3.0​

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Umfanekiso 2. Iphesenti lamaseli e-CD3-CD56+ kumaseli e-NK atholakala e-UCB akhiwe nge-AMMS®NK Cell Culture Kit 3.0​

Izinyathelo zokuzivikela
1, Izidingo zesampula:
① Uma umthamo wegazi lenkaba (okuhlanganisa ne-anticoagulant) ingaphansi kuka-80mL noma isivuno okulindelekile se-NK singaphezulu, kunconywa ukusebenzisa i-HPL esikhundleni se-autologous plasma. Izinga le-cellproliferation ye-HPL cishe liphindwe izikhathi ezingu-1-2 kunalelo le-autologous plasma.
② Ukuhlinzwa kufanele kuqedwe phakathi namahora angu-12 ngemva kokuqoqwa kwegazi.
③ Kunconywa ukuthi ukuminyana okuqandayo kwamasampula afriziwe kube ngu-2-4 x 10 7/mL, futhi izinga lokusebenza ngemva kokutholwa akufanele libe ngaphansi kuka-80%. I-HPL kufanele isetshenziswe esikhundleni se-autologous plasma ngesikhathi kutshalwa amasampula egazi lenkaba. I-Autologous plasma kufanele isetshenziselwe amasampula egazi angaphandle (i-heparin sodium anticoagulant), futhi i-HPL ingasetshenziswa esikhundleni se-autologous plasma.2, Ukuminyana komkhumbi: ukuminyana kwamaseli okuqala ebhodlela kunconywa ukuthi kube amaseli angu-1x10 6/mL. Uma kunamangqamuzana egazi abomvu engeziwe, ukubalwa kwe-fluorescence noma ukubalwa kwe-red blood lysis lysis kufanele kukhethwe ukuze kugwenywe ukuthikameza ukuminyana kwebhodlela lengqamuzana.
3, Ukuminyana kwamanzi: ukuminyana ngemva kokushintshwa koketshezi akufanele kube ngaphansi kuka-1x10 6/mL.
4. Ukusetshenziswa kwe-culture medium:
① I-culture medium kufanele ifudumale kabusha ngokwemvelo ekamelweni lokushisa ngaphambi kokumnika ngakunye.
② Ungalifaki lonke ibhodlela le-culture medium ku-incubator engu-37℃ ukuze uthole ukushisa kabusha, ngaphandle kwalokho kuzosheshisa ukungasebenzi kwama-cytokines endaweni yesiko elingeziwe.
I-GMP Ibanga le-CGT Reagent
③ Isisetshenziswa esiphelele esilungisiwe sineshalofu lesikhathi esifushane, futhi kuyanconywa ukuthi sisetshenziswe cishe isonto elilodwa.
5, Ukuphatha ngendlela efanele kanye nokulondolozwa kwe-plasma: bheka imiyalelo ngemininingwane. I-plasma ye-Centrifuged kufanele icace.
6, Ukusetshenziswa kwezikhwama zesiko: Uma umthamo wamasiko ungaphansi kwe-1L, izikhwama zesiko zidinga ukugoqwa ngaphambi kokubekwa. Kunconywa ukusebenzisa imodeli enconyiwe yenkampani yethu.
7 、 Isikhathi sokufukamela: A, isici kufanele sifukamele u-4 ℃ ngobusuku bonke embhedeni oyisicaba. (Uma kunesimo esiphuthumayo, zama ukufukamela 37℃ amahora ama-2)
8, Ungalinyakazisi ibhodlela lamasiko ngokuthanda kwakho ngesikhathi sokuqala kwesiko: ngaphandle kwalokho, iqembu le-activatedclone lilula ukuntanta phezulu, futhi isici sokumboza sizonciphisa ukusebenza kweqembu lamaseli.
9, Ukusetshenziswa kwezici: Ukuze unciphise ukulahlekelwa kwezinto ezilenga odongeni, kunconywa ukwenza i-centrifugation ngaphambi kokusetshenziswa. Beka ibhodlela eliqukethe izinto kushubhu le-centrifuge elingu-50mL kanye ne-centrifuge ku-1000rpm imizuzu engu-1-2.
10, Ukugcinwa kwemishini: hlola izinga lokushisa kanye nokugxila kwe-CO2 incubator njalo, bese ubuyisela isihlungi ngesikhathi. Ukugcinwa njalo nokuhlanzwa kwekhabhinethi yokuphepha kwezinto eziphilayo.
11, Ukuqapha kwemvelo: buyisela izihlungi eziyinhloko, eziphakathi nendawo neziphezulu njalo ukuze kuqinisekiswe izindinganiso zemvelo zezindawo ezihlanzekile.
12 、 Izinhlobo ezilungisiwe namamodeli wezinto ezisetshenziswayo zokuhlola: umthelela wokushintsha amamodeli kanye nokucaciswa emiphumeleni yokulima kufanele uhlolwe kusenesikhathi, njengamabhodlela amasiko angu-75cm2 nezikhwama zesiko lamaseli.