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发表于 2013-5-21 22:12:51
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shaniu 发表于 2013-5-21 22:00
倪老师，如果要培养分离出来的调节性T细胞，要注意什么啊，成本是不是很可观啊 ...
我没做过体外培养Treg，但从下面这个小鼠Treg的体外培养protocol看，应该成本较贵。
A modified protocol adapted from Tang et al was used to expand regulatory T cells in vitro. Isolated CD4＋CD25＋ regulatory T cells were placed in culture at 2×106 cells/2 mL/well (24 well plate) in RPMI 1640 medium with 7% FBS, 100 U/mL penicillin, 100 g/mL streptomycin, 2 mM glutamine, 50 M 2-ME, 1 mM sodium pyruvate, 0.01 mM nonessential amino acids, and 10 mM HEPES. Anti-mouse CD3– and anti-mouse CD28–coated 4 um polystyrene magnetic beads (Dynal; Invitrogen, Carlsbad, CA) were added at a 1:1 ratio. Recombinant mouse IL-2 (20 ng/mL; R&D Systems, Minneapolis, MN) was added to each well. The cultures were monitored daily and maintained at 0.7 to 1×106 cells/mL. At the end of the seventh day in culture, a magnet was used to remove the beads.
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