Sorting and amplification method of human peripheral blood CD4+CD25+Foxp3+ regulatory T cells
A human peripheral blood, foxp3 technology, applied in the direction of blood/immune system cells, animal cells, vertebrate cells, etc., can solve the problem of small number and so on
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[0027] 1. The materials used in this embodiment are mainly as follows:
[0028] (1) Source of specimens: Peripheral blood specimens were collected from healthy volunteers, a total of 3 cases. Aged 25-32 years old, median age 26 years old, 1 male and 2 females.
[0029] (2) Main reagents: Lymphocyte separation medium was purchased from Sigma, CD3 / CD28 monoclonal antibody-coated Dynalbeads were purchased from Dynal Inc. (Oslo, Norway), CD4+CD25+Treg cell sorting kit, immunomagnetic bead sorting kit AutoMACS Pro Separator, MACS buffer, LD and MS MACS separation columns were purchased from Miltenyi Biotec, Germany. Cell culture reagents: RPMI 1640 cell culture liquid, penicillin and streptomycin double antibodies, fetal bovine serum were purchased from Gibco, flow cytometry antibodies: CD4-FITC, CD25-APC, Foxp3-PE and PE isotype control monoclonal antibodies, fixed Agents and membrane breakers were purchased from BD Pharmigin. The cell counter was Beck-man coulter (USA), and th...
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