Induction method for improving capacity of mesenchymal stem cells in promoting acute myeloid leukemia cell differentiation
A mesenchymal stem cell and cell technology, applied in the field of induction to improve the ability of mesenchymal stem cells to promote the differentiation of acute myeloid leukemia cells, can solve the problems that clinical applications have not been well studied
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Embodiment 1
[0026] Isolation and subculture of umbilical cord mesenchymal stem cells
[0027] 1. Prepare tissue digestion enzymes (type II collagenase 250U / ml, dispase 100U / ml, hyaluronidase 10U / ml), dissolve in α-MEM medium at 37°C, filter and sterilize with a 0.22μm filter for later use ;
[0028] 2. Take the umbilical cord tissue of the newborn baby, put the umbilical cord into a petri dish, wash it with PBS containing 0.1% penicillin-streptomycin double antibody, put it into α-MEM medium, peel off the three blood vessels, and cut the umbilical cord into pieces 1-2mm 3 organization block;
[0029] 3. Mix the shredded tissue pieces with the prepared tissue digestion enzyme solution in a volume ratio of 1:1 in a 50ml centrifuge tube, digest at 37°C, 200rpm for 3 hours, and wait until the tissue pieces are basically completely digested;
[0030] 4. Centrifuge the digested tissue fluid at 300 g for 5 min at 4°C and discard the supernatant. PBS was resuspended in 50ml α-MEM medium, cent...
Embodiment 2
[0034] Optimization of UC-MSCs activation protocol
[0035]1. BMP6 was dissolved in PBS (pH=7.4) at a concentration of 5 μM.
[0036] 2. Add 5 μM biological BMP6 into the complete medium at a ratio of 1:1000 to a final concentration of 5 nM.
[0037] 3. After UC-MSCs grow to 70-80% abundance, add medium containing 5nM BMP6 and culture for 24 hours.
[0038] 4. After activating UC-MSCs for 24 hours, discard the supernatant, wash twice with PBS of pH 7.4, and continue culturing with α-MEM medium containing 10% FBS.
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