Umbilical cord mesenchymal stem cell recovery medium and its preparation method and application
A quality stem cell and culture medium technology, applied in the field of stem cell culture, can solve the problems of secondary crystallization, low cell proliferation rate, and susceptibility to uneven heating, and achieve the effect of improving vitality and proliferation ability.
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Embodiment 1
[0031] The preparation method of the umbilical cord mesenchymal stem cell resuscitation medium provided in this embodiment, the specific scheme is as follows:
[0032] 1. Primary isolation and culture of umbilical cord mesenchymal stem cells
[0033] The umbilical cords of healthy parturients were collected into sterile tubes, and the fresh human umbilical cords were cut into 5cm segments in the ultra-clean workbench, and the residual blood on the surface of the umbilical cords was washed; after the umbilical arteries and veins were stripped, the umbilical cords were placed in double In the anti-DEME medium, cut into tissue pieces of about 3mm; transfer the chopped tissue into a 50mL centrifuge tube, and centrifuge at 300g for 5min; carefully discard the upper layer of medium, add a mixed digestive enzyme equal to the volume of the tissue, and incubate at 37 In a shaker at ℃, shake and digest for 1.5h. The mixed digestive enzymes contain 2g / L type II collagenase, 0.8g / L neutr...
Embodiment 2
[0044] The preparation method of the umbilical cord mesenchymal stem cell resuscitation medium provided in this embodiment, the specific scheme is as follows:
[0045] 1. Primary isolation and culture of umbilical cord mesenchymal stem cells
[0046] The umbilical cords of healthy parturients were collected into sterile tubes, and the fresh human umbilical cords were cut into 5cm pieces in the ultra-clean workbench, and the residual blood on the surface of the umbilical cords was washed; after the umbilical arteries and veins were stripped, the umbilical cords were placed In the anti-DEME medium, cut into tissue pieces of about 3mm; transfer the chopped tissue into a 50mL centrifuge tube, and centrifuge at 300g for 5min; carefully discard the upper layer of medium, add a mixed digestive enzyme equal to the volume of the tissue, and incubate at 37 In a shaker at ℃, shake and digest for 1.5h. The mixed digestive enzymes contain 2g / L type II collagenase, 0.8g / L neutral protease,...
Embodiment 3
[0057] The preparation method of the umbilical cord mesenchymal stem cell resuscitation medium provided in this embodiment, the specific scheme is as follows:
[0058] 1. Primary isolation and culture of umbilical cord mesenchymal stem cells
[0059] The umbilical cords of healthy parturients were collected into sterile tubes, and the fresh human umbilical cords were cut into 5cm segments in the ultra-clean workbench, and the residual blood on the surface of the umbilical cords was washed; after the umbilical arteries and veins were stripped, the umbilical cords were placed in double In the anti-DEME medium, cut into tissue pieces of about 3mm; transfer the chopped tissue into a 50mL centrifuge tube, and centrifuge at 300g for 5min; carefully discard the upper layer of medium, add a mixed digestive enzyme equal to the volume of the tissue, and incubate at 37 In a shaker at ℃, shake and digest for 1.5h. The mixed digestive enzymes contain 2g / L type II collagenase, 0.8g / L neutr...
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