瀵嗗害姊害绂诲績娉曞垎绂?/p>
PBMC
1.
鍦?/p>
15mL
绂诲績绠′腑鍔犲叆
5mL
娣嬪反缁嗚優鍒嗙娑?/p>
Ficoll
銆?/p>
2.
鍙?/p>
5mL
鎶楀嚌闈欒剦琛�涓?/p>
5mL
鏃犺弻
PBS
鎸夌収
1
锛?/p>
1
鍏呭垎娣峰寑锛岀敤绉绘恫绠℃部绠″缂撴參鍙犲姞浜庡垎灞?/p>
娑查潰涓婏紝鍔ㄤ綔涓�瀹氳杞伙紝
娉ㄦ剰淇濇寔娓呮鐨勭晫闈€�傚鍛ㄨ锛?/p>
PBS
锛屾穻宸寸粏鑳炲垎绂绘恫鏈�缁堜綋绉瘮涓?/p>
1
锛?/p>
1
锛?/p>
1
銆?/p>
3.
姘村钩绂诲績
400g
脳
30
鍒嗛挓銆?/p>
锛堟敞鎰忥細涓轰繚璇佸垎绂绘晥鏋滐紝闇�灏嗙蹇冩満鍗囬檷閫熺巼璋冭嚦鏈�浣庯紝鍗冲崌
閫熶负
1
锛岄檷閫熶负
0
锛岃繖鏍疯皟鏁村悗锛屽崌闄嶉�熶細姣旇緝鎱紝鎬讳綋绂诲績鏃堕棿绾︿负
1
灏忔椂銆?/p>
锛?/p>
4.
绂诲績鍚庡彲鐪嬭绠″唴鍒嗕负涓夊眰锛屼笂灞備负琛�娴嗗拰
PBS
锛屼笅灞備富瑕佷负绾㈢粏鑳炲拰绮掔粏鑳烇紝涓眰涓烘穻
宸寸粏鑳炲垎绂绘恫銆?/p>
鍦ㄤ笂銆?/p>
涓眰鐣岄潰澶勬湁涓�浠ュ崟涓牳缁嗚優涓轰富鐨勭櫧鑹蹭簯闆惧眰鐙獎甯?/p>
(
濡備笅鍥?/p>
)
锛?/p>
鍗曚釜鏍哥粏
鑳炲寘鎷穻宸寸粏鑳炲拰鍗曟牳缁嗚優銆傛澶栵紝杩樺惈鏈夎灏忔澘銆?/p>
5.
鍘婚櫎閮ㄥ垎涓婂眰娑蹭綋锛堢敤绉绘恫绠″惛鍙栵紝涓嶅彲鍊惧�掞級
锛屼綑涓嬬害
1mL
锛岀敤绉绘恫鍣ㄦ彃鍒颁簯闆惧眰锛屽惛
鍙栧崟涓牳缁嗚優銆傜疆鍏ュ彟涓�
15mL
绂诲績绠′腑锛屽姞鍏?/p>
5
鍊嶄互涓婁綋绉殑
PBS
锛岀蹇?/p>
300g
脳10
鍒嗛挓锛屾礂娑ょ粏
鑳炰袱娆°�?/p>
6.
鏈绂诲績鍚庯紝寮冧笂娓咃紝鍔犲叆绾㈢粏鑳炶瑙ｆ恫锛屽娓╁鑲?/p>
2
鍒嗛挓锛岃瑙ｇ孩缁嗚優锛屽彲鏍规嵁鎯呭喌閫?/p>
閲忓鍑忔椂闂淬�?/p>
7.
鍔犲叆
10mLPBS
锛岀蹇?/p>
300g
脳10
鍒嗛挓锛屾礂娑ょ粏鑳炰袱娆°�?/p>
8.
鏈绂诲績鍚庯紝寮冧笂娓咃紝鍔犲叆鍚湁
10%FBS
鐨?/p>
RPMI1640
锛岄噸鎮粏鑳烇紝璁℃暟锛岃绠楃粏鑳炴椿鍔涖�?/p>
鐢ㄦ湰娉曞垎绂?/p>
PBMC
锛屾瘡
1mL
鍏ㄨ鍙垎绂诲緱鍒?/p>
1~2
脳10
6
PBMC
锛屼笉鍚屼汉涓綋涔嬮棿瀛樺湪涓�瀹氬樊寮傘�傛椿
缁嗚優鐧惧垎鐜囧湪
95
锛呬互涓娿�?/p>
娉ㄦ剰锛?/p>
琛�娴嗗拰
PBS
娣嬪反缁嗚優鍒?/p>
绂绘恫
绾㈢粏鑳炲拰绮?/p>
缁嗚優
鍗曚釜鏍哥粏
鑳?/p>