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发表于 2013-7-22 15:36:21
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琉璃远 发表于 2013-7-22 16:51
1.细胞的密度，放入DC细胞和淋巴细胞的比例是多少？
2.孵育的时间，3-5天够吗？
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PNAS上的一篇文章应该能够帮助你：
1、T淋巴细胞通常为4×10E5/孔，DC的量可以按照需要不同，可以做梯度。
2、孵育一般5天。
Homeostatic T cell proliferation in a T cell-dendritic cell coculture system
T Cell–DC Coculture. For suspension cultures, CFSE-labeled 2C T cells (usually 400,000/well) were mixed with various numbers of purified DC in 24-well plates in RPMI 1640 medium containing 10% heat-inactivated FCS, 2 mM L-glutamine, 100 units/ml penicillin, and 100 g/ml streptomycin. For reaggregated organ cultures (ROCs), T cells and DC were mixed together in suspensions and then pelleted by centrifugation. After removal of the supernatant, cell pellets were pipetted as a standing drop onto a nucleopore filter placed on a Gelfoam sponge in RPMI 1640 medium with supplements. Cells were cultured at 37° in a humidified 5% CO2 incubator for 5 days before analysis.
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发表于 2013-7-22 15:36:20
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