產品特色
使用血清成分中的Afamin蛋白質將Wnt3a安定化，使其保有高效能Wnt活性
以TOP FLASH熒光素酶測定結果確認我們的商品具有足夠的Wnt3a活性以進行良好的類器官培養
使用能代替血清作用的Afamin蛋白質穩定Wnt3a，可以實現無血清下的類器官培養
由人類大腸所培育的類器官影像
類器官培養常用的Wnt3a在無血清培養基下，會有無法保持良好活性的問題。
而我們的培養液是由含有Wnt3a與血清中能用於穩定Wnt3a的Afamin蛋白質的細胞培養上澄液而來。
所以可以實現無血清下的穩定類器官培養。
本產品用於類器官培養的技術原理
Afamin/Wnt3a CM是將大阪大學蛋白質研究所高木教授與慶應義塾大學醫學部佐藤教授的共同研究成果轉化而成的商品。穩定化後的Wnt3a活性提高、使類器官長期的良好培養化為可能。
2016年Mihara與其研究團隊發現血清成分中的一種Afamin蛋白質和Wnt3a形成複合体後能保持Wnt3a活性，也發現了Afamin/Wnt3a複合体用於類器官的長期培養的可能性。
Afamin使Wnt3a穩定化的機制
2016年Mihara與其研究團隊發現了由血清成分而來的
Afamin蛋白質可以與Wnt3a形成複合體，
使Wnt3a在無血清的類器官培養基中得以穩定。
Afamin/Wnt3a複合體的高活性
透過TOP FLASH熒光素酶測定來進行
Afamin/Wnt3a複合体與重組Wnt3a的活性結果比較、
我們可以看出使用Afamin蛋白質穩定化的Afamin/Wnt3a擁有比較高的活性
使用Afamin/Wnt3a複合体的類器官培養結果
從Afamin/Wnt3a複合體
用於人類大腸類器官的培養、繼代後的結果
與一般市售的重組Wnt3a或含有血清的
Wnt3a Conditioned Medium的培養結果進行比較，
可以發現我們的商品能進行更加長期穩定的培養。
Afamin/Wnt3a CM用於
LGR5+細胞的持續培養
於Lgr5 promoter調節下，我們分別使用Afamin/Wnt3a CM與一般市售的重組Wnt3a進行可表達tdTomato螢光蛋白的類器官培養並比較LGR5+細胞的生長狀況，
實驗結果發現使用Afamin/Wnt3a CM進行培養能高效維持LGR5+細胞的生長。
Afamin/Wnt3a CM
Commercialized recombinant Wnt3a※
Negative Control
基於Nature Protocol使用濃度做設定
與市售Wnt3a的差異
一般類器官培養用的Wnt3a大多來自於市面上販賣的重組Wnt3a或是由ATCC等機構取得可表達Wnt3a的細胞的Conditioned Medium。但由這些方法取得的Wnt3a大多不會進行Wnt3a訊號的活性測試以確認其是否有足夠的效能支持類器官的培養，這也是培養失敗很大的可能原因之一。
此外，如果使用可表達Wnt3a的細胞的Conditioned Medium的話，需要添加血清以維持Wnt3a活性，但我們已經知道血清會對細胞的增殖與培養產生一定影響。所以不須添加血清，並且經過活性測試過具有足夠效能的本商品相信可以為您的類器官培養提供極大幫助。
| Afamin/Wnt3a CM | Recombinant Wnt3a | Wnt3a CM prepared for lab use |
|
| Wnt3a活性 | 藉由Afamin蛋白質使Wnt3a穩定並產生高活性 | 對於LGR5+幹細胞的維持及長期間的類器官培養，所需要的活性不足 | 高活性要藉由血清來維持 |
| 效能評價 | 經過檢測具有類器官培養所需要的良好活性 | 有經過Wnt信號的活性評價測試 | 因為生產批次差的關係，活性的評價可能產生較大誤差 |
| 血清使用 | 無血清 | 無血清 | 因為含有血清所以會影響類器官培養 |
Afamin/Wnt3a CM
Afamin/Wnt3a CM是在無血清培養基中，由共同表達Afamin蛋白質與Wnt3a蛋白質的細胞培養上澄液（Conditioned Medium）而來。所以在無血清培養基中也相信能擁有很好的Wnt3a活性。
Afamin/Wnt3a CM的制备方法
導入Wnt3a及Afamin
基因的細胞
對導入基因的細胞進行大量培養，
這些細胞會在在培養基中分泌Afamin/Wnt3a複合體
培養上澄液(Conditioned Medium)
回収、經滅菌過濾處理後
分裝至小瓶（進行活性評価）
| Afamin/Wnt3a CM 培養液的簡介 | |||||
|---|---|---|---|---|---|
| Code No. | 商品名稱 | 主要成分 | 包装規格 | 溶液 | 有效期限 |
| J2-001 | Afamin/Wnt3a CM | Mouse Wnt3a Human Afamin |
10 mL | Advanced D-MEM/F12 |
記載於標籤上 |
Recombinant Afamin/Wnt3a
Recombinant Afamin/Wnt3a
| Recombinant Afamin/Wnt3a 培養液的簡介 | |||||
|---|---|---|---|---|---|
| Code No. | 商品名稱 | 主要成分 | 包装規格 | 溶媒 | 有效期限 |
| J2-002 | Recombinant Afamin/Wnt3a | Mouse Wnt3a Human Afamin |
60 µg/ 300 µL |
20 mM Tris-HCl (pH 7.4), 150 mM NaCl |
記載於標籤上 |
使用Recombinant Afamin/Wnt3a
進行LGR5陽性細胞的持續培養
我們分別使用Recombinant Afamin/Wnt3a與市售他牌的重組Wnt3a於Lgr5 promoter的調節下進行了可表達tdTomato螢光蛋白的類器官培養，實驗結果發現即使是低濃度的Recombinant Afamin/Wnt3a也可以維持LGR5陽性細胞的良好生長。
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