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Chondrex 胶原诱导的小鼠关节炎(CIA模型)指南

发布者:北京博蕾德生物 发布时间:2024-04-22 分享到:

Chondrex胶原诱导的小鼠关节炎(CIA)实验指南

2023-10-12 16:19   点击次数:476 关键词: Chondrex, CIA,胶原,佐剂,关节炎
 
胶原诱导的小鼠关节炎(CIA)指南
小鼠品系选择:
DBA/1(DBA)小鼠或C57BL/6(B6)小鼠
1)7-8周龄
2)SPF条件
3)高脂饮食

推荐的佐剂和胶原
货号 佐剂   货号 胶原
7002 不完全佐剂IFA(DBA-只在加强免疫用)   20011 鸡II 型胶原,冻干(DBA 或B6)
7008 完全佐剂CFA,1mg/ml(DBA)   20012 鸡II 型胶原,2mg/ml(DBA 或B6)
7001 完全佐剂CFA,4mg/ml(DBA)   20021 牛II 型胶原,冻干(DBA)
7023 完全佐剂CFA,5mg/ml(DBA)   20022 牛II 型胶原,2mg/ml(DBA)

免疫程序(选择其中一种)
方案1:单次免疫(DBA小鼠)
0天:尾根部皮下注射胶原和完全佐剂(4mg/ml)配成的乳剂
0-28天:每7天对关节炎评分
28-56天:每2-3天关节炎评分,关节炎发病率90-100%
56天:实验结束
方案2:加强免疫
0天:尾根部皮下注射胶原和完全佐剂(1mg/ml,DBA 小鼠)或(完全佐剂(5mg/ml,B6小鼠)配成的乳剂
0-28天:每7天对关节炎评分
21天:尾根部皮下注射胶原和不完全佐剂(DBA 小鼠)或完全佐剂(5mg/ml,B6小鼠)配成的乳剂。
28-56天:每2-3天关节炎评分,关节炎发病率90-100%(DBA), 50-70% (B6)
56天:实验结束
用分散机乳剂制备,(不要用2个注射器推送制备)
  1. 准备好实验仪器(图1. 刀头, b.10ml管(3通栓塞),1ml注射器)
  2. 锁上活塞
  3. 置于冰上
  4. 加入完全佐剂
  5. 逐滴加入等体积的胶原溶液
  6. 10000-30000rpm 混匀2min。混匀过程中,上下移动针筒。
  7. 置于冰上5min。
  8. 10000-30000rpm 混匀2min。混匀过程中,上下移动针筒。
  9. 滴入水中检测乳剂混匀质量。
  10. 注射器吸取乳剂
乳剂检测

注射:


a用注射器吸取乳剂(不要有气泡) b针头(25 or 27 gauge x 5/8)         c.固定小鼠装置

皮下免疫乳剂:

免疫后:






注意:一定不要腹腔或静脉免疫小鼠
 

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