メモ

サイクロスポリンA (CSA)（山下潤先生@京大）

内皮細胞への分化誘導を抑えて、心筋分化誘導を促進するらしい。

Biochem Biophys Res Commun. 2009 Jan 30;379(1):115-20. Epub 2008 Dec 16.
Cyclosporin-A potently induces highly cardiogenic progenitors from embryonic stem cells.

Yan P, Nagasawa A, Uosaki H, Sugimoto A, Yamamizu K, Teranishi M, Matsuda H, Matsuoka S, Ikeda T, Komeda M, Sakata R, Yamashita JK.

Laboratory of Stem Cell Differentiation, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.

分化のプロトコールを間違いを指摘されたので、削除。ご指摘を誠にありがとうございます。

Thanks, DK. Below is the memo from your e-mail and google wave.

使用した細胞は

ｍES cell（EMG7） alpha-MHC promter-GFP
Mitomycin C treated OP9 (MMC-OP9) staining with PKH67

EMG7 mouse ES cells that carry mouse a- myosin heavy chain (MHC) promoter-driven EGFP gene were used for this study [8]. OP9 stroma cells were maintained as described [11].

EMG7 on MMC-OP9, or type IV collagen plate (BD) in a-MEM supplemented with 10% FCS
↓ 96 - 108 hrs (medium change/ 2 days)
↓ FACS by Flk1 (+), E-cad (-), PI (-)
Flk1 (+) cells
↓ ↓ on MMC-OP9 staining with PKH67 for 2 days
↓ Flk-2d (FCV progenitor cell) (Flk1 (+), CXCR4 (+), VE-cadherin (-))
↓
↓ on MMC-OP9 staining with PKH67 for 6 days
Flk-6d (Cardiomyocyte)

GFP+ cells were evaluated as differentiated cardiomyocyte.

http://www.fasebj.org/cgi/reprint/04-3540fjev1