Magnetic red blood cell clusters for enriching circulating tumor cells based on magnetic separation method
A magnetic separation and tumor cell technology, applied in the field of biomedicine, can solve the problems of adverse effects on the purity of CTCs, difficulty in removing white blood cells, and affecting the separation purity of CTCs
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Embodiment 1
[0053] The features and advantages of the present invention can be further understood through the following detailed description in conjunction with the accompanying drawings. The examples provided are only illustrative of the method of the present invention and do not limit the rest of the present disclosure in any way. [Example 1] Preparation of normal red blood cell clusters using 5-6 μm magnetic microspheres with carboxyl groups on the surface
[0054] Modification process of red blood cells: Mix 30 μL fresh red blood cells from healthy people with 100 μL PBS solution of DSPE-PEG-FA (1 mg / mL) with a molecular weight of 5000, incubate at 4°C for 30 minutes, centrifuge and wash three times with PBS to obtain folic acid Modified red blood cells. Then mix the erythrocytes with surface-modified folic acid and 10 mg / mL polybrene PBS solution, incubate at 4°C for 30 minutes, wash with PBS for three times to obtain erythrocytes with surface-modified polybrene and folic acid, and ...
Embodiment 2
[0056] The preparation process of fluorescent magnetic red blood cell clusters: first mix 30 μL of fresh red blood cells from healthy people and 1000 μL of DiI (10 μg / mL) in PBS solution, incubate at 37 ° C for 60 minutes, and wash three times with PBS to obtain DiI The stained red blood cells were prepared according to the steps described above to obtain DiI-stained magnetic red blood cell clusters. The fluorescent photos taken under a fluorescent microscope are as follows: image 3 (b) shown. [Example 2] Magnetic red blood cell clusters or fluorescent magnetic red blood cell clusters capture MCF-7 cells
[0057] 1. Magnetic erythrocyte clusters capture MCF-7 cells in PBS: Take 50 μL of magnetic erythrocyte cluster suspension (density 1.2×10 7 cells / mL), add 10uL of MCF-7 cell suspension (density 5×10 5 cells / mL), after mixing evenly, place it at 4°C and incubate for 120min, and blow off the mixture at the bottom of the centrifuge tube after incubation. The bright field pho...
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