Separation method of human peripheral blood mononuclear cells
A separation method, the technology of human peripheral blood, applied in the field of cell separation, can solve the problems of high operation, influence on cell quantity and purity, etc., and achieve the effect of good differentiation ability, high expansion multiple in vitro, high cell viability and purity
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Embodiment 1
[0034] 1. Wipe and disinfect the work surface of the biological safety cabinet with 75% ethanol, and put the anticoagulant tube containing peripheral blood into the ultra-clean bench.
[0035] 2. Transfer the peripheral blood in the anticoagulant tube to a centrifuge tube, and centrifuge at 2000rpm for 10 minutes. After centrifugation, it is divided into two layers, the upper layer is plasma, and the lower layer is blood cell sediment. The upper layer of plasma was collected in another centrifuge tube, marked and placed in a 4°C refrigerator for later use.
[0036] 3. Transfer the blood cells in the lower layer to a 50 mL centrifuge tube, add an equal volume of 6% g / mL hydroxyethyl starch solution, mix well and let it settle naturally for 20 minutes.
[0037] 4. Take another new 50mL centrifuge tube, and add Ficoll lymphocyte separation medium (brand: Axis-shield; supplier: Shenzhen Dakowei Biotechnology Co., Ltd.; article number: AS1114546) to each tube, and after natural sed...
Embodiment 2
[0044] 1. Wipe and disinfect the work surface of the biological safety cabinet with 75% ethanol, and put the anticoagulant tube containing peripheral blood into the ultra-clean bench.
[0045] 2. Transfer the peripheral blood in the anticoagulant tube to a centrifuge tube, and centrifuge at 1500rpm for 8 minutes. After centrifugation, it is divided into two layers, the upper layer is plasma, and the lower layer is blood cell sediment. The upper layer of plasma was collected in another centrifuge tube, marked and placed in a 4°C refrigerator for later use.
[0046] 3. Transfer the blood cells in the lower layer to a 50mL centrifuge tube, add an equal volume of 4% g / mL hydroxyethyl starch solution, mix well and allow it to settle naturally for 15 minutes.
[0047] 4. Take another new 50mL centrifuge tube, and add Ficoll lymphocyte separation medium (brand: Axis-shield; supplier: Shenzhen Dakowei Biotechnology Co., Ltd.; article number: AS1114546) to each tube, and after natural ...
Embodiment 3
[0054] 1. Wipe and disinfect the work surface of the biological safety cabinet with 75% ethanol, and put the anticoagulant tube containing peripheral blood into the ultra-clean bench.
[0055] 2. Transfer the peripheral blood in the anticoagulant tube to a centrifuge tube, and centrifuge at 3000rpm for 12 minutes. After centrifugation, it is divided into two layers, the upper layer is plasma, and the lower layer is blood cell sediment. The upper layer of plasma was collected in another centrifuge tube, marked and placed in a 4°C refrigerator for later use.
[0056] 3. Transfer the blood cells in the lower layer to a 50mL centrifuge tube, add an equal volume of 10% g / mL hydroxyethyl starch solution, mix well and let it settle naturally for 35min.
[0057] 4. Take another new 50mL centrifuge tube, and add Ficoll lymphocyte separation medium (brand: Axis-shield; supplier: Shenzhen Dakowei Biotechnology Co., Ltd.; article number: AS1114546) to each tube, and after natural sediment...
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