Culture method of mesenchymal stem cells contained in umbilical cord blood
A technology of mesenchymal stem cells and culture methods, which is applied in the field of culture of umbilical cord blood containing mesenchymal stem cells, can solve the problem of low success rate of umbilical cord blood mesenchymal stem cell isolation, and achieve risk reduction, immunogenicity, good safety, No effects of exogenous contamination
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Embodiment 1
[0028] A method for culturing umbilical cord blood containing mesenchymal stem cells, specifically comprising the following steps:
[0029] Step S1, collection and transportation of umbilical cord blood; umbilical cord blood comes from fetuses with normal full-term delivery in October 2020. The sample is approved by the hospital ethics committee and consented by the mother and family members before taking the sample. The amount of umbilical cord blood is 90ml; Serum volume fraction is 10%;
[0030] Step S2, extraction of mesenchymal stem cells: put umbilical cord blood and 6% hydroxyethyl starch aqueous solution in a volume ratio of 4:1 into a beaker, stir at room temperature at a speed of 25 r / min for 5 minutes, and then let stand for 30 minutes. Then take the supernatant, centrifuge at 1000r / min for 10min, discard the supernatant, resuspend the cells in the culture medium and add it to Ficoll separation medium containing an equal volume, centrifuge at 1800r / min for 20min, ta...
Embodiment 2
[0040] A method for culturing umbilical cord blood containing mesenchymal stem cells, specifically comprising the following steps:
[0041] Step S1, collection and transportation of umbilical cord blood; umbilical cord blood comes from fetuses with normal full-term delivery in October 2020. The sample is approved by the hospital ethics committee and consented by the mother and family members before taking the sample. The amount of umbilical cord blood is 90ml; Serum volume fraction is 15%;
[0042] Step S2, extraction of mesenchymal stem cells: put umbilical cord blood and 6% hydroxyethyl starch aqueous solution in a volume ratio of 4:1 into a beaker, stir at room temperature at a speed of 28 r / min for 5 minutes, and then let stand for 38 minutes. Then take the supernatant, centrifuge at 1000r / min for 10min, discard the supernatant, resuspend the cells in the culture medium and add it to Ficoll separation medium containing an equal volume, centrifuge at 1800r / min for 20min, ta...
Embodiment 3
[0052] A method for culturing umbilical cord blood containing mesenchymal stem cells, specifically comprising the following steps:
[0053] Step S1, collection and transportation of umbilical cord blood; umbilical cord blood comes from fetuses with normal full-term delivery in October 2020. The sample is approved by the hospital ethics committee and consented by the mother and family members before taking the sample. The amount of umbilical cord blood is 90ml; Serum volume fraction is 20%;
[0054] Step S2, extraction of mesenchymal stem cells: put umbilical cord blood and 6% hydroxyethyl starch aqueous solution in a volume ratio of 4:1 into a beaker, stir at room temperature at a speed of 30 r / min for 5 minutes, and then let stand for 45 minutes. Then take the supernatant, centrifuge at 1000r / min for 10min, discard the supernatant, resuspend the cells in the culture medium and add it to Ficoll separation medium containing an equal volume, centrifuge at 1800r / min for 20min, ta...
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