Positive circulating tumor cell enrichment method
A tumor cell positive technology, applied in the field of positive enrichment of circulating tumor cells, can solve problems such as difficult to ensure specificity and recovery rate at the same time, analysis troubles, affecting the accuracy, sensitivity and specificity of tumor cell detection, to avoid Effects of irreversible damage, maintenance of cell viability, and broadening of utility
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[0057] The first embodiment of the present invention is:
[0058] In order to verify the capture ability of this enrichment method against different cell lines, the MCF-7 and NCI-H226 cell lines pre-labeled by Cell Tracker GreenCMFDA were selected respectively, and they were accurately counted and then spiked into the peripheral blood of healthy volunteers. Enrichment, statistical recovery rate, the specific operation method is as follows:
[0059] 1. PBMC blood separation: Before the experiment, take out the sample density separation solution and equilibrate to room temperature, add 3mL sample density separation solution in the lymphatic separation tube in advance, and then add 3mLPBS and 3mL healthy volunteers' peripheral blood in sequence and add accurate counts Cell Tracker Green CMFDA pre-labeled MCF-7 and NCI-H226 cells, 2000rpm, centrifugation for 20min; use a Pasteur pipette to penetrate the buffy coat layer (mononuclear cell enrichment area) to suck, transfer to a new cent...
Example Embodiment
[0071] The second embodiment of the present invention is:
[0072] In order to verify the possibility of cell culture in vitro after obtaining the target cells using this method, peripheral blood samples of lung cancer patients were used to isolate circulating tumor cells, and relevant technical methods were used to obtain stable proliferation cell lines. The specific operation methods are as follows:
[0073] 1. PBMC blood separation: Before the experiment, take out the sample density separation solution and equilibrate to room temperature, add 3mL sample density separation solution in advance in the lymphatic separation tube, and then add 3mLPBS and 3mL peripheral blood of lung cancer patients in order, and centrifuge at 2000rpm for 20min; use The Pasteur pipette extends into the white membrane layer (mononuclear cell enrichment area) of the liquid surface to suck and transfer to a new centrifuge tube, and discard the red blood cell layer at the bottom of the lymphatic separation ...
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