鐢㈠搧浠嬬垂
绱拌優鍚嶇ū锛�浜虹磪绱拌優鐧借鐥呮穻宸寸窗鑳�
褰㈡厠鐗规€э細娣嬪反姣嶇窗鑳炴ǎ
鐢熼暦鐗规€э細鎳告诞
鐗瑰緛鐗规€э細瑭茬窗鑳炲爆浜�IL-3鍙奊M-CSF渚濊炒鍨�銆�
鍩归姊濅欢锛� 10%FBS
鍌充唬鏂规硶锛� 2.0E+04cells/ml
鍌充唬鎯呮硜锛�鐒′唬娆�
鍑嶅瓨姊濅欢锛� 90%FBS+10%DMSO
鐢㈠搧鍚嶇ū | 浜虹磪绱拌優鐧借鐥呮穻宸寸窗鑳� |
鑻辨枃绨＄ū | TF-1 |
鐙€鎱� | 璨煎 |
绱拌優鍐峰噸淇濆瓨锛�
1.鍑嶅瓨娑诧細92%瀹屽叏鍩归鍩�+8%DMSO锛堝彲浠ユ牴鎿氬椹楀姊濅欢鑷閬告搰锛�
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�
绱拌優鍩归鐨勬搷浣滄椹燂細
1.浜虹磪绱拌優鐧借鐥呮穻宸寸窗鑳�鍚歌蛋鐢ㄤ簬鍩归鍩�锛屽姞鍏�3-5mL PBS鍚庤紩杓曟檭鍕曞煿椁婄摱娼ゆ礂绱拌優锛�
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婄毧浣胯儼閰舵蹈娌掔窗鑳炶〃闈�锛屽煿椁婄摱鏀�37搴﹀煿椁婄娑堝寲锛�
锛堢窗鑳炲皪浜庢秷鍖栨瘮杓冩晱鎰燂紝娑堝寲閬庡害鏈冨毚閲嶅奖闊跨窗鑳炵媭鎱�锛屽皫鑷寸窗鑳炲偝浠ｆ浜℃紓娴垨鑰呭偝浠ｅ悗涓嶉暦銆傜窗鑳炴秷鍖栧埌绱拌優闁撻殭璁婂ぇ浣嗘湭鑴惤锛屽彲浠ヨ紩杓曞惞涓嬫檪鍗冲彲绲傛锛岀姝㈡秷鍖栧埌绱拌優瀹屽叏婕傛诞?锛涢澒铓ｆ瘞楹渞鐩￠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瀹屽叏鍩归鍩虹祩姝㈡秷鍖�锛岃紩杓曞惞涓嬬窗鑳炴贩鍕�锛�
4.灏囨贩鍕荤殑绱拌優1000rpm锛堢磩150g锛夐洟蹇�3min锛屾涓婃竻锛屽啀鐢ㄦ柊楫煿椁婂熀閲嶆嚫37搴﹀煿椁婄绻肩簩鍩归锛�
鍌充唬姣斾緥: 涓嶅悓绱拌優鐢熼暦閫熷害涓嶄竴锛屽叿楂斿偝浠ｆ瘮渚嬭绱拌優鐢熼暦閫熷害鑰屽畾锛屽ぇ閮ㄥ垎绱拌優閬╃敤1:3-1:4鍌充唬锛岀敓闀疯純鎱㈢殑绱拌優鍙互1:2鍌充唬銆�
