Peripheral blood samples from the four groups were collected in heparin anti-coagulated vacutainer tubes (BD Pharmingen, CA, USA). After diluted twice by PBS, the blood was added carefully onto the Ficoll layer (Blood: PBS: Ficoll = 1:1:1) and centrifuged at 800 × g for 30 min with brake off. Mononuclear layer was obtained and washed twice by PBS. The PBMC aliquots were used immediately for Flow Cytometry Staining or stocked frozen in fetal calf serum containing 10% DMSO and 5% Dextran at −80°C until further test.
Heparin anti coagulated vacutainer tubes
Manufactured by BD
Sourced in United States
Heparin anti-coagulated vacutainer tubes are blood collection tubes that contain the anticoagulant heparin. These tubes are designed to collect and preserve blood samples for laboratory analysis.
Automatically generated - may contain errors
4 protocols using heparin anti coagulated vacutainer tubes
1
PBMC Isolation from Heparinized Blood
2
Isolation and Purification of Decidual and Peripheral Blood Gamma Delta T Cells
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Decidual sample isolation was performed according to previous report [34] . Brie y, fresh decidua samples were washed and minced into small pieces. Decidual tissues were released by digesting the tissues with 1 mg/mL collagenase type IV (Sigma, USA) and 150 U/mL of DNase I (Applichem, Germany) in RPMI 1640 medium for 1 hour at 37℃. The suspensions were strained through nylon mesh and then loaded onto a percoll density gradient (60%, 40%, 20%) to purify the lymphocytes. Blood samples were obtained in heparin anti-coagulated vacutainer tubes (BD Biosciences, USA). Peripheral blood mononuclear cells were isolated according to previous report [35] . The DICs and PBMCs were re-suspended in RPMI1640 supplemented with 10% FBS, 1% penicillin and streptomycin, and cultured at 37℃, 5% CO2.
Human γδ T cells isolation γδ T cells were separated from decidua and PBMCs by positively selection using a magnetic isolation kit (Miltenyi Biotec, Germany) according to the manufacturer instructions. The purity of the enriched γδ T cells was evaluated by ow cytometry (Becton Dickinson, USA). The purity of the isolated cells exceeded 90%.
Human γδ T cells isolation γδ T cells were separated from decidua and PBMCs by positively selection using a magnetic isolation kit (Miltenyi Biotec, Germany) according to the manufacturer instructions. The purity of the enriched γδ T cells was evaluated by ow cytometry (Becton Dickinson, USA). The purity of the isolated cells exceeded 90%.
3
Plasma HIV Viral Load Quantification
4
Isolation and Purification of Decidual and Peripheral Blood Gamma Delta T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Decidual sample isolation was performed according to previous report [34] . Brie y, fresh decidua samples were washed and minced into small pieces. Decidual tissues were released by digesting the tissues with 1 mg/mL collagenase type IV (Sigma, USA) and 150 U/mL of DNase I (Applichem, Germany) in RPMI 1640 medium for 1 hour at 37℃. The suspensions were strained through nylon mesh and then loaded onto a percoll density gradient (60%, 40%, 20%) to purify the lymphocytes. Blood samples were obtained in heparin anti-coagulated vacutainer tubes (BD Biosciences, USA). Peripheral blood mononuclear cells were isolated according to previous report [35] . The DICs and PBMCs were re-suspended in RPMI1640 supplemented with 10% FBS, 1% penicillin and streptomycin, and cultured at 37℃, 5% CO2.
Human γδ T cells isolation γδ T cells were separated from decidua and PBMCs by positively selection using a magnetic isolation kit (Miltenyi Biotec, Germany) according to the manufacturer instructions. The purity of the enriched γδ T cells was evaluated by ow cytometry (Becton Dickinson, USA). The purity of the isolated cells exceeded 90%.
Human γδ T cells isolation γδ T cells were separated from decidua and PBMCs by positively selection using a magnetic isolation kit (Miltenyi Biotec, Germany) according to the manufacturer instructions. The purity of the enriched γδ T cells was evaluated by ow cytometry (Becton Dickinson, USA). The purity of the isolated cells exceeded 90%.
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