Method for reversibly separating human adipose mesenchymal stem cells through monoclonal antibody L-NGFR immunomagnetic beads
A technology of immune magnetic beads and mesenchymal stem cells, which is applied in the direction of animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of poor activity, time-consuming and laborious, and low purity of mesenchymal stem cells, and achieve low cost and pain. Small, active effect
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[0024] (1) Preparation of immunomagnetic beads: Co-precipitation method was used to prepare supercis Fe with a particle size of 20±5nm 3 o 4 magnetic nanoparticles.
[0025] Measure an equal volume of 1M FeSO 4 ·7H 2 O solution and 1.75M FeCl 3 solution, mixed rapidly, where [Fe 3+ ]: [Fe 2+ ]=1.75:1; dropwise add 5M NaOH solution to pH=7, at this time, brown particles are formed; then add dropwise 0.5M NaOH solution to pH=11, continue to stir, add absolute ethanol, after standing for 10min, the temperature Raise to 80°C for aging; stir, and add 100 μL, 0.001mol / L surfactant sodium dodecylbenzenesulfonate at the same time; after 30 minutes, use a strong magnet to settle, separate the supernatant, and use distilled water and anhydrous The precipitate was washed repeatedly with ethanol until the pH of the washing water was about 7; the precipitate was placed in a vacuum drying oven and dried at 75°C for 5 hours to obtain super-compliant Fe3O4 nano-magnetic particles.
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