Method for isolating uterine membrane mesenchymal stem cells by dissolving menstrual red blood cells with tea saponin
A technology for red blood cells and stem cells, applied in the biological field, can solve the problems of cell damage, affecting the number of cell cultures, etc., and achieve the effects of good cell activity, simple and controllable components, and convenient operation.
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Embodiment 1
[0027] A method for dissolving menstrual blood red blood cells with tea saponin to separate uterine mesenchymal stem cells, comprising the steps of:
[0028] (1) Reagent preparation: prepare a tea saponin solution, the tea saponin solution is a PBS solution with a tea saponin concentration of 750 mg / L, sterilized by filtration; wherein the tea saponin powder is commercially available.
[0029] (2) Obtain cell pellet: filter 20ml of menstrual blood sample, centrifuge at 300g for 3 minutes to remove the upper layer of plasma, and obtain cell pellet A; add 45ml of tea saponin solution to 5ml of cell pellet A, mix well, let stand for 0.5-2.5min, After mixing evenly, centrifuge at 300 g for 5 min to remove the supernatant and obtain cell pellet B.
[0030] The menstrual blood sample is menstrual blood on the second or third day of the female menstrual cycle, and an equal volume of buffer solution containing at least one antibacterial substance is added, and the antibacterial substa...
Embodiment 2
[0033] A method for dissolving menstrual blood red blood cells with tea saponin to separate uterine mesenchymal stem cells, comprising the steps of:
[0034] (1) Reagent preparation: prepare a tea saponin solution, the tea saponin solution is a PBS solution with a tea saponin concentration of 375 mg / L, sterilized by filtration; wherein the tea saponin powder is commercially available.
[0035] (2) Obtain cell pellet: filter 20ml of menstrual blood sample, centrifuge at 300g for 3 minutes to remove the upper layer of plasma, and obtain cell pellet A; add 45ml of tea saponin solution to 5ml of cell pellet A, mix well, let stand for 0.5-2.5min, After mixing evenly, centrifuge at 300 g for 5 min to remove the supernatant and obtain cell pellet B.
[0036] The menstrual blood sample is menstrual blood on the second or third day of the female menstrual cycle, and an equal volume of buffer solution containing at least one antibacterial substance is added, and the antibacterial substa...
Embodiment 3
[0039] A method for dissolving menstrual blood red blood cells with tea saponin to separate uterine mesenchymal stem cells, comprising the steps of:
[0040](1) Reagent preparation: prepare a tea saponin solution, which is a PBS solution with a tea saponin concentration of 1500 mg / L, sterilized by filtration; wherein the tea saponin powder is commercially available.
[0041] (2) Obtain cell pellet: filter 20ml of menstrual blood sample, centrifuge at 300g for 3 minutes to remove the upper layer of plasma, and obtain cell pellet A; add 45ml of tea saponin solution to 5ml of cell pellet A, mix well, let stand for 0.5-2.5min, After mixing evenly, centrifuge at 300 g for 5 min to remove the supernatant and obtain cell pellet B.
[0042] The menstrual blood sample is menstrual blood on the second or third day of the female menstrual cycle, and an equal volume of buffer solution containing at least one antibacterial substance is added, and the antibacterial substance is cefotaxime s...
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