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绱�(x矛)鑳炲悕绋憋細浜洪鎴愮簴缍窗(x矛)鑳炵郴
褰㈡厠(t脿i)鐗规€э細鎴愮簴缍窗(x矛)鑳炴ǎ銆€
鐢熼暦鐗规€э細璨煎鐢熼暦
鐗瑰緛鐗规€э細瑭茬窗(x矛)鑳炲爆灏堝埄淇濊棌锛屽叾鐗瑰緛鐗规€э細灏氭湭鍏枊銆�
鍩归(y菐ng)姊濅欢锛� MEM-EBSS锛�
Minimum Essential Medium (MEM Eagles with Earle锛噑 Balanced Salts) 10%FBS
鍌充唬鏂规硶锛� 1锛�
3鍌充唬锛�3-4澶╁偝1娆�
鍌充唬鎯呮硜锛� C8
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� | 浜洪鎴愮簴缍窗(x矛)鑳炵郴 |
鑻辨枃绨$ū | CNLMG-B5538SKIN |
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﹂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.浜洪鎴愮簴缍窗(x矛)鑳炵郴鍚歌蛋鐢ㄤ簬鍩归(y菐ng)鍩�锛屽姞鍏�3-5mL PBS鍚庤紩杓曟檭鍕曞煿椁�(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�锛�
锛堢窗(x矛)鑳炲皪浜庢秷鍖栨瘮杓冩晱鎰燂紝娑堝寲閬庡害鏈冨毚(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鍗冲彲绲傛锛岀姝㈡秷鍖栧埌绱�(x矛)鑳炲畬鍏ㄦ紓娴�銆傛贩鍕荤窗(x矛)鑳炴檪鐩¢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�锛�
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�