绉嶅睘锛氬皬榧燑/span>

缁嗚優绯荤壒寰侊細The CATH.a cell line was established from cultures of a tumor that arose in the brain of a transgenic mouse銆侟/span>

The transgene contain the SV40 T antigen coding sequence linked to a 783 bp portion of the rat tyrosine hydroxylase gene.

The cells express tyrosine hydroxylase and dopamine beta-hydroxylase, and contain neurofilaments.

缁勭粐鏉ユ簮锛氳剳

鐢熼暱鐗规€э細鎴愬洟鍗婃偓娴拰璐村娣峰悎鐢熼暱

寰敓鐗╋紝鏀師浣撴娴嬶細闃存€?/span>

瀹夊叏鎬э細鎵€鏈夎偪鐦ゅ拰鐥呮瘨杞煋鐨勭粏鑳炲潎瑙嗕负鏈夋綔鍦ㄧ殑鐢熺墿鍗卞鎬э紝蹇呴』鍦ㄤ簩绾х敓鐗╁畨鍏ㄥ彴鍐呮搷浣滐紝骞惰娉ㄦ剰闃叉姢锛屾墍鏈夊簾娑插強鎺ヨЕ杩囨缁嗚優鐨勫櫒鐨块渶楂樺帇鐏弻鍚庢柟鑳戒涪寮冦€侟/span>

鍩瑰吇鏉′欢锛欬/span>

瀹屽叏鍩瑰吇鍩猴細鍚?mML-璋锋皑閰拌兒RPMI1640锛?8%锛涢┈琛€娓?%锛涜儙鐗涜娓?%銆侟/span>

鍩瑰吇娓╁害锛?7鈩冿紱CO2锛?%銆侟/span>

浼犱唬鏂规硶锛欬/span>

鏀跺埌缁嗚優鍚庯紝鍦ㄥ€掔疆鏄惧井闀滀笅瑙傚療鏁翠釜缁嗚優鐢熼暱鎯呭喌銆備箣鍚庣敤75%閰掔簿鍠锋磼鏁翠釜鐡舵秷姣掑悗鏀惧埌瓒呰弻鍙板唴锛屼弗鏍兼棤鑿屾搷浣滐紝鎵撳紑缁嗚優鍩瑰吇鐡讹紝鍚稿嚭鍩瑰吇娑插埌鏃犺弻鐨勭蹇冪鍐咃紝1000杞?鍒嗙蹇?0鍒嗛挓鍚庡純涓婃竻锛屽姞鏂伴厤鐨勫畬鍏ㄥ煿鍏绘恫10ml鎺ョ鍒板煿鍏荤摱鍐呭煿鍏?濡傛灉缁嗚優澶氾紝涔熷彲鎺ョ涓ょ摱)銆傚師鐡舵崲10ml鏂伴矞鍩瑰吇娑插悗缁х画鍩瑰吇銆侟/span>

缁嗚優浼犱唬鏃讹紝鍙杞绘媿鍩瑰吇鐡跺悗鍔犲鍩瑰吇娑蹭竴鍒嗕负浜屽嵆鍙€傚鏋滄湁浜涚粏鑳炴媿鎵撲笉涓嬫潵锛屽彲鎸変互涓嬫柟娉曞仛锛欬/span>

1.鍚稿嚭鎷嶆墦涓嬬殑缁嗚優鎮恫锛岀敤涓嶅惈閽欍€侀晛绂诲瓙鐨凱BS娲?-2娆°€侟/span>

2. 鍔?ml娑堝寲娑?0.25%Trypsin-0.53mM EDTA)浜庡煿鍏荤摱涓紝鍊掕浆鏀句簬37鈩冨煿鍏荤1-3鍒嗛挓棰勭儹锛岀劧鍚庡皢鍩瑰吇鐡剁炕杩囨潵璁╄儼閰朵笌缁嗚優闈㈡帴瑙﹀ぇ绾?0-30绉掑悗锛屽湪鍊掔疆鏄惧井闀滀笅瑙傚療缁嗚優娑堝寲鎯呭喌锛岃嫢缁嗚優澶ч儴鍒嗗彉鍦嗭紝杩呴€熸嬁鍥炴搷浣滃彴锛岃交鏁插嚑涓嬪煿鍏荤摱鍚庡姞6-8ml/鐡跺畬鍏ㄥ煿鍏诲熀鍗冲彲銆侟/span>

娉細1銆佽瀵熺粏鑳?*鍦ㄤ綆鍊嶉暅(4鎴?X鐗╅暅)涓嬭繘琛岋紝鍚﹀垯涓嶈兘鍑嗙‘鍒ゆ柇缁嗚優鐨勪紶浠ｅ瘑搴︺€傜湅缁嗚優鐨勫舰鎬佽鍦?0X鐗╅暅涓嬨€侟/span>

2銆佺摱涓繍杈撳煿鍏诲熀涓嶈兘閲嶅鍐嶇敤,璇锋崲鐢ㄥ姞鍙屾姉鐨勬柊鍩瑰吇鍩猴紝缁嗚優鍐诲瓨鍚?鍩瑰吇鍩轰腑鍙笉鍔犱换浣曟姉鐢熺礌銆侟/span>

3銆佹湁浜涚粏鑳炶创澹佷笉鐗紝濡傚彂鐜拌创澹佺粏鑳炴湁鑴辫惤锛屽彲绂诲績鍚规墦鍚庢帴绉嶅埌鏂扮摱鍐呫€侟/span>

4銆佹敹鍒扮粏鑳炲悗锛岃嫢鍙戠幇鍩瑰吇鐡剁牬鎹熴€佹湁娑叉孩鍑哄強缁嗚優鏈夋薄鏌擄紝璇峰強鏃朵笌鎴戜滑鑱旂郴銆侟/span>

鍐诲瓨鏂规硶锛欬/span>

鍐诲瓨娑诧細90%瀹屽叏鍩瑰吇娑叉垨鐗涜娓咃紝10%DMSO 銆傜幇鐢ㄧ幇閰嶃€侟/span>

鍌ㄥ瓨锛氭恫姘偍瀛樸€侟/span>