鐢㈠搧浠嬬垂
绱拌優鍚嶇ū锛�浜鸿儙鍏掕兏鑵虹窗鑳炴牚
褰㈡厠(t脿i)鐗规€э細
鐢熼暦鐗规€э細璨煎鐢熼暦銆€
鐗瑰緛鐗规€э細
鍩归(y菐ng)姊濅欢锛� DMEM-H锛�
Dulbecco锛噑 Modified Eagle锛噑 Medium (DME H-21 4.5g/Liter Glucose) 10%FBS
鍌充唬鏂规硶锛�
鍌充唬鎯呮硜锛�涓嶈┏
鍑嶅瓨姊濅欢锛�鍩虹鍩归(y菐ng)鍩�+5%DMSO+20%FBS銆€
鐢㈠搧鍚嶇ū | 浜鸿儙鍏掕兏鑵虹窗鑳炴牚 |
鑻辨枃绨$ū | HFT-8810[HFT8810] |
鐙€鎱�(t脿i) | 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(y菐ng)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閬庡鍚庢恫姘繚瀛樸€�
绱拌優鍩归(y菐ng)鐨勬搷浣滄椹燂細
1.浜鸿儙鍏掕兏鑵虹窗鑳炴牚鍚歌蛋鐢ㄤ簬鍩归(y菐ng)鍩猴紝鍔犲叆3-5mL PBS鍚庤紩杓曟檭鍕曞煿椁�(y菐ng)鐡舵饯娲楃窗鑳烇紱
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拌優琛ㄩ潰锛屽煿椁�(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�(t脿i)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銆傛贩鍕荤窗鑳炴檪鐩¢噺杓曟煍锛屼笉瑕佸惞鍑哄ぇ閲忔埃娉°€傦級
3.鍔犲叆6-8ml瀹屽叏鍩归(y菐ng)鍩虹祩姝㈡秷鍖栵紝杓曡紩鍚逛笅绱拌優娣峰嫽锛�
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�(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�