I-NK kunye ne-Til yeSeli yeSeli yeSoftwers (K562)
I-NK kunye ne-Til yeSeli yeSeli yeSoftwers (K562)
✔ Ngaphambi - Ukudityaniswa, i-Multi - i-cytokine iphuculwe i-NK / til yokwandiswa.
✔ Iqinisekisiwe kuwo omabini intambo yegazi kunye nemithombo ye-PBMC - Imoto - ilungele.
✔ Ngaphambi - bakufanelekele ukungqubana okungaphakathi ngedatha epheleleyo.
Amagqabantshintshi:
I I-NK / TIL ERGER ine Iinjineli ze-K562 ye-K562 yeeseli zondla Ukuveza il - Ama-21 kunye nezinye iiCytokines eziphambili. Xa idityaniswa nemithombo yeendaba emiselweyo yesosi, le nkqubo yenza ukuba:
-
Ukuphakama - Ukwanda kokuphumelela ye Iiseli zeNK kunye neeTils kwii-PBMCs okanye igazi -
Iphezulu - Ukucoceka kwabantu (> Iipesenti ezingama-90 zeepesenti)
-
Ukukhawuleza, ukuSebenzisa Phantsi kwe-cytoskine esebenzayo
Izicelo eziphambili
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Ukusebenza kunye nokwandiswa ye Iiseli ze-NK, Imoto - i-NKS, kunye neMils
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Inkqubo yokuphuhliswa kwenkqubo: Kudala - Ukuphuculwa kwenqanaba lokuhamba kweeseli zeseli
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Ukwenza imveliso ye-GPP: Inkxaso I-Ind - Ukwenza izifundo kwaye Iposi - Inkqubo yokuvunywa itshintsha
Ulawulo kunye neziBonelelo zentsebenzo
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I-GMP - ehambelana Ukuqulunqwa
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Iyahambelana, iphakamileyo - Ukwandiswa kwesivuno (ixhomekeke ku I-1000 - Songa)
-
Ngaphambi - Iqinisekisiwe ukusetyenziswa kweklinikhi
Ishiti yedatha
I-NK yeseli yokwandiswa kunye nokusebenza kwi-Renonors ezininzi
I-Donor - I - Ukwahlukano kwe-Doors kwi-NK, ukucoceka kweseli
Iziphumo zokuphucula intsholongwane e - I-hancerc kwi-lentivivral yogcino kwiiseli zeNK
I-NK yokuTshintshela kweeSeli: I-Viral E - i-hancerc iphumza kukhuphiswano kwi-MOI 1 iPaneli yasekhohlo: i-cytometry ye-Il - I-15 ye-Cytometry ye-Il - I-NK ye-NK ye-NK - i-hancerc
Iphaneli yasekunene: Ukuhamba kwe-cytometry ye-Il - Iiseli ezili-15 ze-NK ngaphandle kokutshintshela
I-Viral E - i-hancerc vs. Abakhuphisana: T - Utshintshiselwano lweseli kwi-Moi 5 ngasekhohlo: I-Cytometry yemoto - I-T.
Ekunene: Ukuhamba kwe-cytometry yemoto - Iiseli ze-T ngaphandle kokutshintshela
Iziphumo zokuphucula intsholongwane e - I-hancer A + i-villa e - i-hancer D indibaniselwano kwi-lentiviviral edluliselweyo kwii-HSCs
Uhlalutyo oluthelekisayo lwe-HSC
| Inombolo yesiriyeli | Igama le-regentand | I-Dosage (ye ~ 1 - 2e6 pbmc yokuqalisa) |
| 1 | I-Serum - I-PreeIum Free | 2 l |
| 2 | Il - 2 | Ii-Stick ezi-4 (ezingama-200,000 IU / intonga) |
| 3 | I-0.9% yenaliti | ~ 500 ml |
| 4 | I-20% ye-isbum yomntu i-albumin | ~ 15 ml |
| 5 | I-Ficoll Lymphocyte yedwa | ~ 25 ml |
| 6 | I-NC - Igumbi le-200 legumbi (okanye elingana) | Njengoko kufuneka |
| 7 | 6 - iplate | 1 |
| 8 | I-TOSK | 1 |
| 9 | I-T75 FLAASK | 1 |
| 10 | I-T175 flask | 2 |
| 11 | Ingxowa yenkcubeko engama-640 cm² | 2 |
| 12 | I-15 ml centrifuge tube | 1 |
| 13 | I-1.5 Ml Centrifuge Tube | Njengoko kufuneka |
| 14 | I-10 ml / 25 ml pipettes | Njengoko kufuneka |
| 15 | I-1 ml / 200μ μl ye-pipette pipette | Njengoko kufuneka |
Ukucaciswa
| Ipharamitha yokusebenza | Ukucaciswa |
| I-NK / Imoto - I-NK yokwanda | > Iintsuku ezingama-5 000 (iintsuku ezili-14) |
| Ukusebenza kweseli | ≥90% |
| Ukucoceka kweseli (CD56⁺ / CD3⁻) | > I-95% |
Ulwazi lokuhambisa
Sinikezela ngothutho oluhlaziyiweyo kuyo yonke imiyalelo. Ngokwesiqhelo, iodolo yakho iya kufika ngaphakathi kwe-5 - Iintsuku ezisi-7 zeshishini eUnited States kwaye kwiintsuku ezili-10 zomsebenzi wamazinye amazwe. Nangona kunjalo, nceda uqaphele ukuba ukuhanjiswa kweendawo ezisemaphandleni kunokuthatha ixesha elide.
Ixesha lokuhambisa: Ii-odolo zihlala zisetyenzwa ngaphakathi 1 - iintsuku ezintathu zomsebenzi. Nje ukuba i-odolo yakho ithunyelwe, uyakufumana i-imeyile yokuqinisekisa ngolwazi lokulandela umkhondo.
Ulwazi olubalulekileyo
Ukuqhubekeka kocwangco: Emva kokuba i-odolo ihlawulwe, i-Warehouse yethu idinga ixesha lokuqhuba iodolo yakho. Uya kufumana isaziso emva kokuba i-odolo yakho ithunyelwe.
Amaxesha okuhambisa: Kwiimeko ezininzi, iphakheji iya kusiwa ngexesha eliqikelelweyo lokufika. Nangona kunjalo, umhla wokwenyani unokuchaphazeleka ngamalungiselelo eenqwelomoya, iimeko zemozulu, kunye nezinye izinto zangaphandle. Ixesha lokuhanjiswa kwexesha liya kuba ngaphezulu kwesiqhelo semiyalelo ebandakanya ukhuseleko okanye izinto ezenziwe ngokwezifiso. Nceda ujonge ulwazi lokulandelela ngowona mhla uchanekileyo wokuhanjiswa.
Imicimbi yokuhambisa: Ukuba ufumanisa ukuba iphakheji yakho ayihanjiswanga ngexesha elimiselweyo; Ulwazi lokulandela umkhondo lubonisa ukuba iphakheji ihanjisiwe kodwa awukayifumani; Okanye iphakheji yakho ibandakanya izinto ezilahlekileyo okanye ezingachanekanga okanye eminye imiba yempahla, nceda uqhagamshelane nenkonzo yethu yabathengi kwiintsuku ezisi-7 zomhla wokuhlawula ukuze sikwazi ukujongana nale micimbi ngokukhawuleza.
Ukulandelela umkhondo
Nje ukuba i-odolo yakho ithunyelwe, uyakufumana i-imeyile ngenombolo yokulandela umkhondo kunye nekhonkco lokulandelela ukuthunyelwa kwakho.
Unokulandelela i-odolo yakho ngqo kwiwebhusayithi yethu ngokungena kwiakhawunti yakho kwaye ujonge imbali yakho ye-oda.
Izithintelo zokuhambisa
Nceda ugcwalise idilesi yesitrato ngokweenkcukacha, hayi ibhokisi ye-PO okanye idilesi yomkhosi (AP). Ngaphandle koko, kuya kufuneka sisebenzise i-EMS yokuhanjiswa (ihamba kancinci kunabanye, ukuthatha malunga 1 - iiyure ezi-2 okanye ixesha elide).
Imisebenzi yesiko kunye nomgaqo-nkqubo werhafu
Nceda uqaphele ukuba nayiphi na imisebenzi yesiko, irhafu, okanye intlawulo yokungenisa okwenziweyo ngexesha lokuhambisa luxanduva lomthengi. Ezi ntlawulo ziyahluka ngokuxhomekeke kwilizwe eliya kulo kwaye zimiselwa ngabasemagunyeni basekhaya.
Ngokuthenga kwiwebhusayithi yethu, uyavuma ukuhlawula nayiphi na imisebenzi esebenzayo okanye irhafu enxulumene ne-oda yakho. Asinaxanduva lokulibaziseka okubangelwa kukucocwa kwesiko.
Umgaqo-nkqubo wePakeji yephakheji
Nje ukuba i-odolo yakho ifikile kwindawo echongiweyo yePickup okanye indawo yokuHanjiswa, nceda uqinisekise ukuqokelelwa ngokukhawuleza. Ukuba iphakheji ayithanga ngexesha elichongiweyo, siya kuthumela isikhumbuzo nge-imeyile okanye nge-SMS. Nangona kunjalo, ukuba iphakheji ayiqokelelwa ngaphakathi kwexesha elichaziweyo, kwaye nayiphi na ilahleko okanye umonakalo ovela ngenxa yoko, umthengi uya kuba noxanduva. Ngobubele kukhumbuza ukuba uqokelele iphakheji yakho ngokukhawuleza ukuphepha nayiphi na imiba enokubakho.
Qaphela: Njengoko imveliso yethu iwela phantsi kodidi olukhethekileyo, ibuyise kunye nokubuyiselwa kwemali ayamkelwa.
