鐢�(ch菐n)鍝佷粙绱�
绱�(x矛)鑳炲悕绋憋細浜烘€ユ€�T绱�(x矛)鑳炵櫧琛€鐥呯窗(x矛)鑳�
褰㈡厠(t脿i)鐗规€э細娣嬪反姣嶇窗(x矛)鑳炴ǎ
鐢熼暦鐗规€э細鎳告诞鐢熼暦
鐗瑰緛鐗规€э細瑭茬窗(x矛)鑳炵敘(ch菐n)鐗╃偤IL-2锛岃綁(zhu菐n)鍏ョ啋鍏夌礌閰跺熀鍥�锛孴绱�(x矛)鑳炲彈楂旓紝鎶楀師琛ㄩ仈(d谩)CD3銆�
鍩归(y菐ng)姊濅欢锛� RPMI 1640 (w/o Hepes) 10%FBS銆€
鍌充唬鏂规硶锛�
鍌充唬鎯呮硜锛�
鍑嶅瓨姊濅欢锛�鍩虹(ch菙)鍩归(y菐ng)鍩�+5%DMSO+20%FBS銆€
鐢�(ch菐n)鍝佸悕绋� | 浜烘€ユ€绱�(x矛)鑳炵櫧琛€鐥呯窗(x矛)鑳� |
鑻辨枃绨$ū | JurkatCA |
鐙€鎱�(t脿i) | 璨煎 |
绱�(x矛)鑳炲喎鍑嶄繚瀛橈細
1.鍑嶅瓨娑诧細92%瀹屽叏鍩归(y菐ng)鍩�+8%DMSO锛堝彲浠ユ牴鎿�(j霉)瀵�(sh铆)椹�(y脿n)瀹ゆ浠惰嚜琛岄伕鎿囷級
2.闄嶆韩姝ラ锛�4搴�10min锛�-20搴�2h锛�-80閬庡鍚庢恫姘繚瀛�銆�
绱�(x矛)鑳炲煿椁�(y菐ng)鐨勬搷浣滄椹燂細
1.浜烘€ユ€�T绱�(x矛)鑳炵櫧琛€鐥呯窗(x矛)鑳�鍚歌蛋鐢ㄤ簬鍩归(y菐ng)鍩猴紝鍔犲叆3-5mL PBS鍚庤紩杓曟檭鍕�(d貌ng)鍩归(y菐ng)鐡舵饯娲楃窗(x矛)鑳�锛�
2.鍚稿共鍑圥BS鍚庡姞鍏�1mL 0.25%鑳伴叾-0.53mM EDTA锛岃紩杓曟悥鏅冨煿椁�(y菐ng)鐨夸娇鑳伴叾娴告矑绱�(x矛)鑳炶〃闈�锛屽煿椁�(y菐ng)鐡舵斁37搴﹀煿椁�(y菐ng)绠辨秷鍖栵紱
锛堢窗(x矛)鑳炲皪(du矛)浜庢秷鍖栨瘮杓冩晱鎰�锛屾秷鍖栭亷搴︽渻(hu矛)鍤�(y谩n)閲嶅奖闊跨窗(x矛)鑳炵媭鎱�(t脿i)锛屽皫(d菐o)鑷寸窗(x矛)鑳炲偝浠f浜℃紓娴垨鑰呭偝浠e悗涓嶉暦銆傜窗(x矛)鑳炴秷鍖栧埌绱�(x矛)鑳為枔闅欒畩澶т絾鏈劔钀�锛屽彲浠ヨ紩杓曞惞涓嬫檪(sh铆)鍗冲彲绲傛锛岀姝㈡秷鍖栧埌绱�(x矛)鑳炲畬鍏ㄦ紓娴�?锛涢澒铓�?x矛)鑳炴檪(sh铆)鐩¢噺杓曟煍锛屼笉瑕佸惞鍑哄ぇ閲忔埃娉�銆傦級
3.鍔犲叆6-8ml瀹屽叏鍩归(y菐ng)鍩虹祩姝㈡秷鍖�锛岃紩杓曞惞涓嬬窗(x矛)鑳炴贩鍕伙紱
4.灏囨贩鍕荤殑绱�(x矛)鑳�1000rpm锛堢磩150g锛夐洟蹇�3min锛屾涓婃竻锛屽啀鐢ㄦ柊楫煿椁�(y菐ng)鍩洪噸鎳�37搴﹀煿椁�(y菐ng)绠辩辜绾�(x霉)鍩归(y菐ng)锛�
鍌充唬姣斾緥: 涓嶅悓绱�(x矛)鑳炵敓闀烽€熷害涓嶄竴锛屽叿楂斿偝浠f瘮渚嬭绱�(x矛)鑳炵敓闀烽€熷害鑰屽畾锛屽ぇ閮ㄥ垎绱�(x矛)鑳為仼鐢�1:3-1:4鍌充唬锛岀敓闀疯純鎱㈢殑绱�(x矛)鑳炲彲浠�1:2鍌充唬銆�