Human primary T cells were isolated from fresh or frozen peripheral blood obtained from healthy donors at the Hematology Department of Hospital Universitario Reina Sofía (Córdoba, Spain). All donors gave their written informed consent, and the study was performed according to the guidelines of the Regional Government Junta de Andalucía-Consejería de Salud ethics committee and requirements under the Spanish specific regulation (RD 1301/2006) and International Conference of Good Clinical Practice. Blood was diluted 1/2 to 1/4 in PBS, and mononuclear cells were collected after Ficoll separation (Lymphosep, Biowest). Cells were washed twice at 200 g for 10 min, and then, Pan-T cells were isolated following MACS Express Separator (Miltenyi Biotec) protocol and then seeded at 106 cells/ml in TexMACS supplemented with interleukin-2 (IL-2) (20 IU/mL, Miltenyi Biotec). T cells were activated with TransAct (Miltenyi Biotec) during 48 h before lentiviral transduction following the manufacturer’s recommendations.
Macs express separator
Manufactured by Miltenyi Biotec
Sourced in Germany
The MACS Express Separator is a compact and user-friendly magnetic separation device designed for easy and efficient cell separation. It utilizes magnetic labeling technology to quickly and effectively isolate target cells from complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Texmacs, by Miltenyi Biotec (2 mentions)
Lymphosep, by Biowest (1 mentions)
Interleukin 2 (il 2), by Miltenyi Biotec (1 mentions)
Pan t cell isolation kit, by Miltenyi Biotec (1 mentions)
T cell transact, by Miltenyi Biotec (1 mentions)
Il 7 and il 15, by Miltenyi Biotec (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
2 protocols using macs express separator
1
Isolation and Activation of Primary Human T Cells
2
Primary T Cell Isolation and Activation
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Primary T cells were isolated from fresh or frozen apheresis products obtained from healthy donors at the Hematology Department of the Hospital Universitario Reina Sofía (Córdoba, Spain). All donors gave their written informed consent, and the study was performed according to the guidelines of the local ethics committee and complies with the requirements regarding quality and safety for donation, obtaining, storage, distribution, and preservation of human cells and tissues under the Spanish-specific regulation (RD-L 9/2014). Pan-T cells were isolated by negative selection using immunomagnetic beads (Pan T cell Isolation Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) and following MACSExpress Separator (Miltenyi Biotec) or AutoMACs Pro Separator’s (Miltenyi Biotec) protocol and cultured in TexMACS (Miltenyi Biotec) supplemented with 20 UI/mL of IL-2 (Miltenyi Biotec) for EGFP experiments or with 10 ng/mL of IL-7 and IL-15 (Miltenyi Biotec) for CAR experiments. T cells were activated with T cell TransAct (Miltenyi Biotec) during 48 h as recommended by the manufacturer.
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