A kind of hemizygous capri cell preparation method
A hemizygous and cell technology, applied in the field of preparation of hemizygous CAPRI cells, can solve the problems of poor constitution, hinder the implementation of CAPRI cell treatment plan, and cannot achieve collection, etc. Effects of proliferation and activation ability
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Embodiment 1
[0027] Example 1: The patient, 60 years old, female, after the modified radical mastectomy for stage I breast cancer, decreased the number of white blood cells due to chemotherapy, and was unable to withstand the process of collecting mononuclear cells by a blood cell separator because of poor physical condition, chose the hemizygous CAPRI cell preparation method of the present invention. The present invention will be described in detail below through examples.
[0028] Take 100ml of peripheral blood from the patient and 150ml from the hemizygous donor, add heparin sodium with a final concentration of 1.5×105U / L for anticoagulant treatment, and divide into 50ml centrifuge tubes according to the volume of peripheral blood obtained, and centrifuge at 1600rpm for 20- After 30 minutes, the supernatant was discarded, and 25ml of normal saline was added to the centrifuge tubes, carefully mixed, and slowly distributed along the tube wall into 15ml centrifuge tubes pre-added with 5ml o...
Embodiment 2
[0033] Example 2: Patient, male, 54 years old, lung squamous cell carcinoma with bone metastases, white blood cells < 4000 / ml after radiotherapy + chemotherapy, not suitable for collecting mononuclear cells with a blood cell separator, choose the hemizygous CAPRI of the present invention Cell preparation method. The present invention will be described in detail below through examples.
[0034] Take 120ml of peripheral blood from the patient and 150ml of peripheral blood from the hemizygous donor, add heparin sodium with a final concentration of 1.5×105U / L for anticoagulant treatment, and divide into 50ml centrifuge tubes according to the volume of peripheral blood obtained, and centrifuge at 1800rpm for 20- After 30 minutes, the supernatant was discarded, and 25ml of normal saline was added to the centrifuge tubes, carefully mixed, and slowly distributed along the tube wall into 15ml centrifuge tubes pre-added with 5ml of Ficoll-Hypaque lymphocyte separation medium. After cent...
Embodiment 3
[0039] Example 3: CAPRI cell toxicity test in vitro.
[0040]The present invention also detects the in vitro cytotoxicity of the obtained hemizygous CAPRI, the in vitro cytotoxicity of the CAPRI cells (CD3mAb-CAPRI) stimulated by CD3mAb alone and the CAPRI cells (CD3 / CD133mAb-CAPRI) co-stimulated by CD3mAb and CD133mAb For comparison, the specific experimental steps are as follows: CAPRI cells harvested from two different stimulation methods were used as effector cells, and the human lung squamous cell carcinoma NCI-H520 cell line in the logarithmic growth phase was stained with 0.4% trypan blue and counted As target cells, use serum-free 1640 medium to adjust the cell concentration of effector cells and target cells to 1×106 / ml with different effector-target ratios of 2.5:1, 5:1, 10:1, and 20:1, and add them to 96-well plates , 100 μl / well, in addition, set blank control wells, sample control wells, and sample maximum enzyme activity control wells, set 3 duplicate wells for e...
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