A method for preparing induced pluripotent stem cell chromosomes
A technology of pluripotent stem cells and chromosomes, which is applied in the field of preparation of induced pluripotent stem cell chromosomes, can solve the problems of optimal final concentration and optimal blocking time interval without reference data, chromosomes affecting karyotype analysis, high-resolution chromosome length, etc. , to achieve the effect of high success rate, large quantity and high resolution
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Embodiment 1
[0043] A method for preparing induced pluripotent stem cell chromosomes, comprising the steps of:
[0044] (1) Cultivation of stem cells: Human stem cells were placed at 37°C with 5% CO 2 Open culture in the incubator, the medium (commercially available) was changed daily;
[0045] (2) Synchronization of the cell cycle: replace the fresh medium at 8:00 a.m. the day before stem cell harvesting, add 100 microliters of fluorouracil deoxynucleoside (concentration 1 μg / ml) to the culture flask at 10:00 a.m., and continue to add Cultivate in an incubator; add 200µl thymine (concentration 2.4mg / ml) to the culture flask at 8:00 the next day, and continue culturing, add 100µl colchicine (concentration 20µg / ml) to the culture flask at 11:00 am, and mix well After placing in the incubator for 15 minutes, take out the culture bottle for harvesting stem cells;
[0046] (3) Stem cell harvesting:
[0047] A. Aspirate the culture medium in the culture bottle into the centrifuge tube, add 2...
Embodiment 2
[0064] A method for preparing induced pluripotent stem cell chromosomes, comprising the steps of:
[0065] (1) Stem cell culture: Stem cells were placed at 37°C, 5% CO 2 Open culture in the incubator, the medium (commercially available) was changed daily;
[0066] (2) Synchronization of the cell cycle: replace the fresh medium at 8:00 a.m. the day before stem cell harvesting, add 100 microliters of fluorouracil deoxynucleoside (concentration 1 μg / ml) to the culture flask at 10:00 a.m., and continue to add Cultivate in an incubator; add 200µl thymine (concentration 2.4mg / ml) to the culture flask at 8:00 the next day, and continue culturing, add 100µl colchicine (concentration 20µg / ml) to the culture flask at 11:00 am, and mix well After placing in the incubator for 15 minutes, take out the culture bottle for harvesting stem cells;
[0067] (3) Stem cell harvesting:
[0068] A. Aspirate the culture medium in the culture bottle into the centrifuge tube, add 2ml of normal salin...
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