iProtocol yoMfuniselo
I-cytotoxicity % ibalwa ngale equation ingezantsi.
I-Cytotoxicity % = (Amanani aphilayo olawulo - Amanani aphilileyo aphathwayo) / Amanani okulawula okuphilayo × 100
Ngokubhala iiseli zethumba ekujoliswe kuzo nge-non-toxic, non-radioactive calcein AM okanye i-transfect nge-GFP, sinokubeka iliso ekubulaweni kweeseli zethumba ngeeseli ze-CAR-T.Ngelixa iiseli zomhlaza ezijoliswe kuzo ziya kubhalwa nge-calcein eluhlaza i-AM okanye i-GFP, iiseli ezifileyo azikwazi ukugcina idayi eluhlaza.Hoechst 33342 isetyenziselwa ibala zonke iiseli (zombini iiseli T kunye neeseli ithumba), ngenye indlela, iiseli ithumba ekujoliswe unako ukungcoliswa inwebu ebotshwe calcein AM, PI isetyenziselwa ibala iiseli ezifileyo (zombini iiseli T kunye neeseli ithumba).Esi sicwangciso sokungcolisa sivumela ukucalulwa kweeseli ezahlukeneyo.
E: T ixhomekeke kwi-Cytotoxicity ye-K562
Umzekelo Hoechst 33342, CFSE, PI imifanekiso fluorescent ziiseli K562 ekujoliswe t = 3 iiyure
Iziphumo zemifanekiso yefluorescent ibonise ukonyuka kweHoechst+CFSE+PI+ Iiseli ekujoliswe kuzo njengo-E: umlinganiselo we-T wonyukile
