Method for detecting lymphocyte transformation by utilizing medicament cytochalasins B
A technology of lymphocyte transformation and cytochalasin, which is applied in the field of medical testing, can solve the problems of limited laboratory research, high laboratory requirements, and limited wide application, and achieve the effect of low equipment requirements, low experimental cost, and accurate results
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[0027] (1) Materials.
[0028] 1 Phytohemagglutinin (Sigma Company), cytochalasin B (Sigma Company).
[0029] 2 The culture medium is RPMI1640 (PAA), containing 10% calf serum (Sijiqing), and 1% double antibody (Geno).
[0030] 3 Heparin anticoagulation tubes.
[0031] 4 Normal saline, 087% NH 4 Cl solution, Giemsa staining solution.
[0032] (2) Operation method.
[0033] 1. Sterile blood collection, 0.5ml each, divided into 2Gy irradiation group (immunosuppressive group) and non-irradiation group, respectively injected into 4.5ml RPMI1640 culture medium containing PHA, the final concentration of PHA was 50μg / ml, mixed well, placed in Incubate in a 37°C incubator for 2 days.
[0034] 2. On the third day, cytochalasin B was added to the culture medium with a final concentration of 3 μg / ml, and the cells were cultured for 24 hours.
[0035] 3. Lyse red blood cells: Discard the supernatant on day 4, add 0.87% NH prewarmed at 37°C 4 Cl solution 3ml, incubate in a 37°C i...
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