求助

未来之星

Co-culture of MSCs and T lymphocytes. The co-culture of the MSCs and T cells was carried out using 24-well plates (contact culture) and the Transwell system (isolated culture) in which the T cells and MSCs were physically separated by a membrane permeable for soluble factors. The inner hanging cell culture insets with 0.4‑μm pore size membrane of a 24-well plate were purchased from Millipore (Billerica, MA, USA). The BM-, AT-, WJ- and PL-MSCs were seeded at 105 cells/well in regular 24-well plates and 24-well Transwell plates containing α‑MEM, 10% FBS and 100 U/ml penicillin/streptomycin. After 24 h, 10 μg/ml mitomycin C (MMC; Sigma) were added to inhibit MSC proliferation, and the cells were incubated for 2 h at 37˚C followed by 5 extensive washes with medium. A total of 105 T cells/well was added and stimulated with 10 g/ml phytohemagglutinin (PHA; Sigma) and 10 ng/ml interleukin (IL)-2 (Sigma). IL-2/PHA-activated T cells were cultured in the presence or absence of MSCs.
5 v# V8 f( g* F( n! b读了这段文字感觉脑子里还没有概念，是不是将MSC作为饲养层后用丝裂霉素处理，然后再将T细胞接种上。一种是直接接种在MSC上（就是文字所说的contact culture），另一种是将T细胞接种到一个网里，然后将网放在已接种MSC的孔里（isolated culture）？不知道文字中所说的24-well plates (contact culture) and the Transwell system (isolated culture) 是不是图中这种？望告知，谢谢2 v" I  B+ W* N: z1 g6 E
24孔培养板链接：http://www.emdmillipore.com/CN/zh/product/Millicell-插入式细胞培养皿%2C-12 mm%2C-聚碳酸酯%2C-0.4 μm,MM_NF-PIHP01250

未来之星

回复 未来之星 的帖子
9 E0 J6 e1 m: w7 N+ e- Q
) y8 }7 T/ S3 v+ R1 W7 q! f为什么不能直接将T细胞消化下来计数，非要用Brdu呢？

倒腾细胞

你这个是直接search的protocol吗？还是文献里的method

未来之星

回复 倒腾细胞 的帖子% w6 p: \6 C" L% b; U( F( @6 c. z

" @0 M8 T9 D" P  c1 s4 C一篇别人文章里面的method