1
瀹為獙涓�
鍔ㄧ墿缁嗚優鐨勪紶浠ｅ煿鍏?/p>
涓�銆佸疄楠岀洰鐨?/p>
1
锛庣啛鎮夌粏鑳炲煿鍏昏繃绋嬩腑鐨勬棤鑿屾搷浣滄妧鏈�?/p>
2
锛庝簡瑙ｇ粏鑳炰紶浠ｅ煿鍏荤殑鍩烘湰鏂规硶鍜屼富瑕佹搷浣滄楠?/p>
浜屻�佸疄楠屽師鐞?/p>
缁嗚優鍩瑰吇
(cell
culture)
鏄寚浠庢満浣撳唴鍙栧嚭鏌愮缁勭粐鎴栫粏鑳烇紝妯℃嫙鏈轰綋鍐呯殑鐢熺悊
鏉′欢浣垮叾鍦ㄤ綋澶栫敓瀛樸�佺敓闀垮拰绻佹畺鐨勮繃绋嬨�傜粏鑳炵殑浣撳鍩瑰吇鍦ㄧ粏鑳炵敓鐗╁鍜屽尰瀛?/p>
鐮旂┒棰嗗煙鏈夌潃鏋佷负骞挎硾鐨勭敤閫旓紝杩欎竴鎶�鏈凡鎴愪负鐮旂┒缁嗚優鐢熺悊銆佺粏鑳炲娈栥�佺粏鑳?/p>
閬椾紶銆佺粏鑳炵檶鍙樺拰缁嗚優宸ョ▼绛夎棰樼殑涓�椤逛笉鍙己灏戠殑鎵嬫銆傜粏鑳炲煿鍏绘妧鏈殑绐佸嚭
浼樼偣鍦ㄤ簬鑳戒负鐮旂┒鑰呮彁渚涘ぇ閲忕殑鐢熺墿鎬х姸鐩稿悓鐨勭粏鑳炰綔涓虹爺绌跺璞★紝渚夸簬浜轰滑鍦?/p>
浣撳鍒╃敤鍚勭涓嶅悓鐨勬柟娉曚粠涓嶅悓鐨勮搴︾爺绌剁粏鑳炵敓鍛芥椿鍔ㄧ殑瑙勫緥銆傚彟澶栵紝鍒╃敤缁?/p>
鑳炲煿鍏绘妧鏈繕鍙娇浜轰滑杈冧负鏂逛究鍦扮爺绌跺悇绉嶇墿鐞嗐�佸寲瀛﹀拰鐢熺墿鍥犵礌瀵圭粏鑳炵粨鏋勫拰
鍔熻兘鐨勫奖鍝嶃�?/p>
缁嗚優鍩瑰吇鍙垎涓哄師浠ｅ煿鍏诲拰浼犱唬鍩瑰吇
2
绉嶆儏鍐点�?/p>
鎵�璋?/p>
鍘熶唬鍩瑰吇
(primary culture)
鏄寚鐩存帴浠庢満浣撳彇鍑虹粍缁囨垨缁嗚優鍚庢墍杩涜鐨勯娆″煿鍏汇�?/p>
鑰?/p>
浼犱唬鍩瑰吇
(subculture)
鏄寚
褰撳師浠ｅ煿鍏荤殑缁嗚優澧炴畺鍒颁竴瀹氬瘑搴﹀悗锛屽皢鍏朵粠鍘熷煿鍏诲鍣ㄤ腑鍙栧嚭锛屼互
1
锛?/p>
2
鎴栧叾浠?/p>
姣斾緥杞Щ鍒板彟涓�涓垨鍑犱釜瀹瑰櫒涓墍杩涜鐨勫啀鍩瑰吇銆備紶浠ｅ煿鍏诲彲绠�绉颁紶浠ｃ�傚湪浣撳
鍩瑰吇杩囩▼涓紝瑕佷娇缁嗚優鑳芥甯稿湴鐢熼暱銆佺箒娈栵紝闇�缁忓父瀵瑰叾杩涜浼犱唬銆備紶浠ｇ殑绱Н
娆℃暟灏辨槸缁嗚優鐨勪唬鏁般�?/p>
鍦ㄤ粠浜嬬粏鑳炲煿鍏诲伐浣滄椂甯稿父浼氭帴瑙﹀埌
缁嗚優绯?/p>
(cell
line)
鍜岀粏鑳炴牚
(cell
strain)
杩?/p>
涓や釜瀹规槗娣锋穯鐨勬蹇点�備竴鑸涓猴紝缁嗚優绯绘寚閫氳繃鍘熶唬鍩瑰吇骞剁粡浼犱唬鍚庢墍褰㈡垚鐨勭粏
鑳炵兢浣擄紝鐢变簬缁嗚優绯绘潵婧愪簬鍘熶唬鍩瑰吇锛岃�屽師浠ｅ煿鍏荤墿涓墍鍚殑缁嗚優绉嶇被杈冨锛屾晠
涓�涓粏鑳炵郴寰�寰�鐢卞涓敓鐗╁鎬х姸涓嶅悓鐨勭粏鑳炵兢浣撴墍缁勬垚锛岃�岀粏鑳炴牚鏄埄鐢?/p>
鍗曠粏
鑳炲垎绂诲煿鍏绘硶鎴栧厠闅嗗舰鎴愭硶
浠庡師浠ｅ煿鍏荤墿鎴栫粏鑳炵郴涓�夋嫨鍑虹殑缁嗚優缇や綋锛屼竴涓粏
鑳炴牚寰�寰�鍏锋湁鐗规畩鐨勭敓鐗╁鎬х姸鎴栨爣璁板苟鍙寔缁瓨鍦ㄣ�?/p>
缁嗚優鍩瑰吇鏄竴绉嶇▼搴忓鏉傘�佹潯浠惰緝澶氫笖瑕佹眰涓ユ牸鐨勫疄楠屾�у伐浣溿�傜敱浜庣粏鑳炲湪
浣撳鐨勭敓闀裤�佺箒娈栦細鍙楀埌娓╁害銆佽惀鍏荤墿璐ㄣ�侀吀纰卞害銆佹笚閫忓帇鍙婂井鐢熺墿绛夊绉嶅洜绱?/p>
鐨勬樉钁楀奖鍝嶏紝鏁呯粏鑳炲煿鍏诲伐浣滅殑鍚勪釜鐜妭濡傚煿鍏诲櫒鐨挎竻娲楁秷姣掋�佽惀鍏绘恫閰嶅埗鍜岄櫎
鑿屻�?/p>
pH
鍊艰皟鏁淬�佹俯搴﹁皟鑺傜瓑鎿嶄綔閮芥湁涓ユ牸鐨勮姹傚拰瑙勫畾锛岀壒鍒娉ㄦ剰鏃犺弻鎿嶄綔锛?/p>
杩欐槸缁嗚優浣撳鍩瑰吇鎴愯触鐨勫叧閿�?/p>
涓夈�佸疄楠屾潗鏂?/p>
浜鸿倽鐧岀粏鑳炴牚
HepG2
鍥涖�佸疄楠屽櫒鏉?/p>
CO
2
鎭掓俯鍩瑰吇绠便�佽秴鍑�宸ヤ綔鍙般�佹亽娓╂按娴寸銆佸�掔疆鐩稿樊鏄惧井闀溿�佺數鐑珮鍘嬭捀姹?/p>
娑堟瘨閿呫�佺粏鑳炲煿鍏荤摱銆佸煿鍏荤毧銆佸惛绠°�侀櫎鑿屾护鍣ㄣ�佸皬鐑ф澂銆?/p>
100mL
鐜荤拑璇曞墏鐡躲�?/p>
姗＄毊鐡跺銆侀厭绮剧伅銆佽鍙风瑪銆?/p>
浜斻�佸疄楠岃瘯鍓?/p>
1
锛?/p>
DMEM
鍩瑰吇娑诧細鐢ㄥぉ骞崇О鍙?/p>
DMEM
鍩瑰吇鍩虹矇鍓?/p>
(
璐ㄩ噺渚濆疄楠屾墍闇�鍩瑰吇娑茬殑鎬?/p>
閲忚�屽畾
)
鍊掑叆鐑ф澂涓紝鎸夎鏄庝功瑕佹眰鍔犲叆涓夎捀姘淬�?/p>
NaHCO
3
銆佽胺姘ㄩ叞鑳猴紝鍒╃敤纾佸姏
鎼呮媽鍣ㄤ娇鍔犲叆鐨勫浐浣撶墿瀹屽叏婧惰В锛?/p>
鐒跺悗鍔犲叆閫傞噺娴撳害涓?/p>
20000
鍗曚綅
/mL
鐨勯潚闇夌礌鍜?