Ekhaya » Kuba CAR-T Cell Therapy

Kuba CAR-T Cell Therapy

  • 1.Ingqokelela
  • 2.Ukwahluka
  • 3.Ukuguqulwa
  • 4.Ukwandiswa
  • 5.Ukuvuna
  • 6.Imveliso ye-QC
  • 7.Unyango

Into esinokuyenza

  • AO/PI Viability
  • Iseli Cytotoxicity
  • UFanelo loTshintsho
  • Iseli Apoptosis
  • Umjikelo weSeli
  • Isiphawuli seCD
  • Iiseli ezonakeleyo
  • Ukubala kweeseli
  • Umgca weSeli
AO/PI Viability
AO/PI Viability

I-Dual-fluorescence Viability (AO/PI), i-Acridine orange (AO) kunye nepropidium iodide (PI) zii-nucleic staining zenucleic kunye ne-acid-binding dayi.I-AO inokungena kwi-membrane yazo zombini iiseli ezifileyo neziphilayo kwaye ingcolise i-nucleus, ivelise i-fluorescence eluhlaza.Ngokwahlukileyo, i-PI inokungena kuphela kwiinwebu eziqhekekayo zeeseli ze-nucleated ezifileyo, zivelise i-fluorescence ebomvu.Itekhnoloji esekwe kumfanekiso we-Countstar Rigel ayibandakanyi amaqhekeza eeseli, inkunkuma, kunye namasuntswana e-artifact kunye neziganeko ezingaphantsi kweeplatelet, ezinika iziphumo ezichanekileyo.Ukuqukumbela, inkqubo ye-Countstar Rigel ingasetyenziselwa inyathelo ngalinye lenkqubo yokwenziwa kweeseli.

Cell Cytotoxicity
Iseli Cytotoxicity

I-T / NK Cell-Mediated Cytotoxicity, Kunyango lweeseli ze-CAR-T ezivunyiweyo ze-FDA zakutshanje, i-T-lymphocyte eyenziwe ngofuzo ibopha ngokuthe ngqo kwiiseli zomhlaza ezijoliswe kuzo (T) kwaye zibabulale.Abahlalutyi be-Countstar Rigel bayakwazi ukuhlalutya le nkqubo epheleleyo ye-T / NK Cell-Mediated Cytotoxicity.

Uphononongo lwe-Cytotoxicity lwenziwa ngokubhala iiseli zomhlaza ekujoliswe kuzo nge-CFSE okanye ngokuzidlulisela nge-GFP.I-Hoechst 33342 ingasetyenziselwa ukungcolisa zonke iiseli (zombini iiseli ze-T kunye neeseli zethumba).Kungenjalo, iiseli zethumba ekujoliswe kuzo zinokudyojwa ngeCFSE.Ipropidium iodide (PI) isetyenziselwa ukudyobha iiseli ezifileyo (zombini iiseli T kunye neeseli zethumba).Ucalucalulo phakathi kweeseli ezahlukeneyo lunokufumaneka kusetyenziswa olu buchule bokungcolisa.

Transfection Efficiency
UFanelo loTshintsho

I-GFP Transfection Efficiency, Kwi-molecular genetics, imodeli eyahlukeneyo yezinto eziphilayo, kunye ne-cell biology, i-GFP gene isetyenziswa rhoqo njengentatheli kwizifundo zokuvakalisa.Okwangoku, izazinzulu zihlala zisebenzisa ii-microscopes ezikhanyayo okanye ii-cytometers ezijikelezayo ukuhlalutya ukuhanjiswa kakuhle kweeseli ezanyisayo.Kodwa ukuphatha itekhnoloji entsonkothileyo ye-cytometer yokuhamba phambili kufuna umntu onamava kwaye oqeqeshwe kakhulu.I-Countstar Rigel yenza abasebenzisi bakwazi ukwenza lula kwaye ngokuchanekileyo uvavanyo olusebenzayo lokutshintshela ngaphandle kweendleko zokusebenza kunye nokugcinwa kweendleko ezinxulumene ne-cytometry yendabuko.

Cell Apoptosis
Iseli Apoptosis

I-Cell Apoptosis, Inkqubela phambili ye-apoptosis yeseli inokujongwa kusetyenziswa i-FITC i-Annexin-V edibeneyo kunye ne-7-ADD.Iintsalela ze-Phosphatidylserine (PS) zihlala zibekwe kwicala langaphakathi le-plasma membrane yeeseli eziphilileyo.Ngexesha le-apoptosis yokuqala, ingqibelelo ye-membrane iyalahleka kwaye i-PS iya kudluliselwa ngaphandle kwenwebu yeseli.Isihlomelo V sinobudlelwane obuluqilima kwi-PS kwaye ke ngoko sesona siphawuli sifanelekileyo kwiiseli zokuqala ze-apoptotic.

Cell Cycle
Umjikelo weSeli

Umjikelo weSeli, Ngexesha lokwahlulwahlulwa kweeseli, iiseli ziqulethe izixa ezandisiweyo zeDNA.Ibhalwe yi-PI, ukunyuka kwe-fluorescence intensity ihambelana ngokuthe ngqo nokuqokelela kwe-DNA.Umahluko intensity fluorescence iiseli enye zezalathi ubume yokwenene umjikelo weseli MCF 7 iiseli baphathwa 4μM of Nocodazole ukubamba ezi seli kwizigaba ezahlukeneyo umjikelo wabo cell.Imifanekiso yentsimi eqaqambileyo efunyenwe ngexesha lovavanyo lwemeko isivumela ukuba sichonge iseli nganye.I-PI fluorescence channel ye-Countstar Rigel ichonga imiqondiso ye-DNA yeeseli enye nakwii-aggregates.Uhlalutyo oluneenkcukacha lwe-fluorescence intensity lunokwenziwa kusetyenziswa i-FCS.

CD Marker
Isiphawuli seCD

I-CD Marker Phenotyping, Iimodeli ze-Countstar Rigel zibonelela ngendlela ekhawulezayo, elula kwaye enovakalelo ngakumbi, i-immuno-based phenotyping yeeseli ngokufanelekileyo.Ngemifanekiso enesisombululo esiphezulu kunye nekhono elinamandla lokuhlalutya idatha ehlanganisiweyo, i-Countstar Rigel ivumela abasebenzisi ukuba bafezekise iziphumo ezithembekileyo ngaphandle kwesidingo sezicwangciso ezibanzi zolawulo olunzima kunye nohlengahlengiso lwembuyekezo ye-fluorescence.

I-Cytokine Induced Killer (CIK) i-cell differentiation ibonisa umgangatho obalaseleyo wokusebenza kwe-Countstar Rigel analyzer ngokuthelekisa ngokuthe ngqo kwii-cytometers ezihamba phambili zeklasi.Ii-PBMC zemouse kwinkcubeko zihlanjululwe nge-CD3-FITC, i-CD4-PE, i-CD8-PE, kunye ne-CD56-PE, kwaye iqhutywe yi-Interleukin (IL) 6. Emva koko yahlalutya ngaxeshanye kunye ne-Countstar® Rigel kunye ne-Flow Cytometry.Kolu vavanyo, i-CD3-CD4 , i-CD3-CD8, ne-CD3-CD56 zahlulahlulwe yangamaqela amathathu, ukujonga umyinge weeseli ezahlukeneyo.

Degenerated Cells
Iiseli ezonakeleyo

Ukufunyanwa kweeSeli eziNcinciweyo nge-Immunofluorescence, ii-antibodies ze-Monoclonal ezivelisa imigca yeeseli ziya kulahlekelwa ezinye ii-clones ezilungileyo ngexesha lokunyuka kweeseli kunye nokudlula ngenxa yokuthotywa okanye ukuguqulwa kofuzo.Ilahleko ephezulu iya kuchaphazela kakhulu imveliso yenkqubo yokuvelisa.Ukubeka iliso kokuthotywa kudlala indima ebalulekileyo kulawulo lwenkqubo yokutshintsha isivuno se-antibodies ukuya kweyona nto iphezulu.

Uninzi lwezilwa-buhlungu ezenziwe kushishino lwe-BioPharma zinokubonwa ngokuleyibhelishwa kwe-immunofluorescence kwaye zihlalutywe ngokobungakanani luchungechunge lwe-Countstar Rigel.Imifanekiso yetshaneli ekhanyayo kunye ne-fluorescence engezantsi ibonisa ngokucacileyo loo ma-clones aphulukene neempawu zawo zokuvelisa izilwa-buhlungu ezifunwayo.Uhlalutyo oluneenkcukacha ngakumbi kunye nesoftware yeDeNovo FCS Express yoMfanekiso iqinisekisa, ukuba i-86.35% yazo zonke iiseli zivakalisa i-immunoglobulins, kuphela yi-3.34 % ecacileyo ngokucacileyo.

Cell Counting
Ukubala kweeseli

I-Trypan (i-Capitalize B ngeBlue) Ukubalwa kweeSeli, i-Trypan blue staining isasetyenziswa kuninzi lweelebhu zenkcubeko yeeseli.

I-Trypan Blue Viability kunye neCell Density BioApp inokufakwa kuzo zonke iimodeli ze-Countstar Rigel.Ii-algorithms zethu zokuqaphela umfanekiso okhuselweyo zihlalutya ngaphezulu kweeparamitha ezingama-20 ukuze zihlele into nganye efunyenweyo.

Cell Line
Umgca weSeli

I-Cell Line Storage QC, Ekugcinweni kweeseli, ingqikelelo yolawulo lomgangatho ophucukileyo iqinisekisa ukhuseleko, ukubekwa kweliso ngokufanelekileyo kuzo zonke iimveliso zeselula.Oku kuqinisekisa umgangatho ozinzileyo weseli cryopreserved, cryo-igcinelwe imifuniselo, uphuhliso lwenkqubo, kunye nemveliso.

I-Countstar Rigel ifumana imifanekiso enesisombululo esiphezulu, ihlalutya iimpawu ezahlukeneyo ze-morphological zezinto zeselula ezifana nobubanzi, imilo, kunye nokuthambekela kokudibanisa.Imifanekiso yamanyathelo ahlukeneyo enkqubo inokuthelekiswa ngokulula komnye nomnye.Ke ukwahluka kobume kunye nokudibanisa kunokubonwa ngokulula, ngokunqanda imilinganiselo yabantu.Kwaye i-database ye-Countstar Rigel inenkqubo yokulawula eyinkimbinkimbi yokugcina kunye nokubuyiswa kwemifanekiso kunye nedatha.

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