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各位大神 小女子最近刚刚接触细胞实验 想问个问题 各位帮帮忙呀 感激不尽~ Short-term TCLs were incubated with irradiated autologous PBMCs and the allergens/peptides of interest.After 48 h, supernatant was collected and tritium-labeled thymidine was added to determine proliferation. The following day,cells were harvested and thymidine uptake was measured with a betaplate counter. Cytokine concentrations (IL-13, IFNγ and IL-10) in the supernatant were determined with ELISA.Forlongterm TCLs, irradiated EBV-transformed B cells were pre-incubated overnight with the allergens/peptides of interest. The next day,the long-term TCLs were added and incubated for another 24 h.Subsequently, thymidine uptake was determined as described above. To determine the degree of proliferation, the stimulation index (SI), i.e. the ratio between the proliferation of allergen/peptide-stimulated cells and non-stimulated cells, was calculated. SI≥2 was considered positive. The major allergens, namely CMP [a mixture of whey (Lactalis, Laval, France) and casein(FrieslandCampina Domo, Amersfoort, The Netherlands) in a 1:1 ratio], casein, whey, β-lactoglobulin and α-LAC (both fromNIZO Food Research BV, Ede, The Netherlands) were tested in a concentration of 50 μg/ml 这段话里面短期和长期培养TCLs是什么意思啊 有什么区别么 在细胞培养中2者有各自适用的情况么 |
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