T/NK Cell Mediated Cytotoxicity Analysis ndi CFSE, Hoechest 33342 ndi PI

Experimental Protocol

cytotoxicity% imawerengedwa ndi equation yomwe ili pansipa.
Cytotoxicity% = (Ziwerengero zowongolera - Ziwerengero zaposachedwa) / Ziwerengero zowongolera × 100
Polemba ma cell a chotupa omwe ali ndi calcein AM yopanda poizoni, yopanda ma radio kapena transfect ndi GFP, titha kuyang'anira kuphedwa kwa ma cell a chotupacho ndi ma cell a CAR-T.Ngakhale maselo a khansa omwe akufuna kukhala ndi moyo adzalembedwa ndi green calcein AM kapena GFP, maselo akufa sangathe kusunga utoto wobiriwira.Hoechst 33342 imagwiritsidwa ntchito pakudetsa ma cell onse (ma cell a T ndi cell chotupa), m'malo mwake, ma cell chandamale amatha kuipitsidwa ndi nembanemba womangidwa calcein AM, PI imagwiritsidwa ntchito pakudetsa ma cell akufa (ma cell a T ndi ma cell chotupa).Njira yodetsa iyi imalola kusankhana kwa ma cell osiyanasiyana.

E: T Retio yodalira Cytotoxicity ya K562

Chitsanzo Hoechst 33342, CFSE, PI fulorosenti zithunzi ndi K562 chandamale maselo pa t = 3 maola
Zithunzi zotsatiridwa ndi fulorosenti zinawonetsa kuwonjezeka kwa ma cell a Hoechst+CFSE+PI+ Target monga E: T chiŵerengero chawonjezeka.