Heparinized whole blood (100 ml) was obtained by venipuncture from healthy human donors, diluted 1:1 in endotoxin-free sterile saline, and separated into components by gradient centrifugation using lymphocyte separation medium (MP Biomedicals, Solon, OH, USA). Mononuclear cells were collected, washed three times with RPMI 1640, and the monocytes were isolated through negative selection using magnetic Dynabeads according to the manufacturer’s protocol (Invitrogen/Life Technologies). Monocytes were cultured in RPMI supplemented with 20% human AB+ serum (Fisher Scientific) and Pen/Strep at 37°C and 5% CO2. After a 4-day maturation period, media was replaced and human monocyte-derived macrophages (HMDM) were harvested for assays on day 7. The human Jurkat T cell line was purchased from ATCC (Manassas, VA, USA) and cultured in RPMI supplemented with 10% heat-inactivated FBS and Pen/Strep. All procedures were approved by the Des Moines University Institutional Review Board before initiation, and all procedures performed were in accordance with the approved protocol.
Human jurkat t cell line
Manufactured by American Type Culture Collection
Sourced in United States
The Human Jurkat T cell line is a well-characterized immortalized T lymphocyte cell line. It is commonly used as a model for T cell signaling and activation studies.
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2 protocols using human jurkat t cell line
1
Isolation and Cultivation of Human Monocyte-Derived Macrophages
2
Isolation and Culture of T Cells
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Blood samples were collected into tubes containing EDTA. Ficoll-Hypaque density gradient centrifugation was used to isolate PBMCs at 300g and 4°C for 10 min. Then magnetic beads coated with anti-human CD3 (Miltenyi Biotec, GmbH, Gladbach, Germany) was used to purify the T cells. The purified T cells were then cultured in Roswell Park Memorial Institute (RPMI) 1640 medium at 37°C in 5% CO 2 with 10% fetal bovine serum, 100 U/mL penicillin, and 100 U/mL streptomycin.
The human Jurkat T cell line (ATCC, Manassas, VA, USA) was cultured in the same condition as the isolated T cells.
The human Jurkat T cell line (ATCC, Manassas, VA, USA) was cultured in the same condition as the isolated T cells.
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