Method for using umbilical stalk placenta to prepare mesenchyma stem cell
A technology of stem cells and placenta, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of potential danger in treatment, potential toxicity of streptomycin to human body, etc., and achieve sensitization and animal origin. Low content of sexual substances, good proliferation potential, effect provided in large quantities
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Embodiment 1
[0026] Example 1: Preparation method of umbilical cord-derived mesenchymal stem cells
[0027] 1. Take the umbilical cord of a healthy full-term baby and wash the surface with saline solution containing gentamicin (concentration 20,000-40,000 U / L) and heparin (concentration 50-200 U / ml) to remove residual blood and possible bacterial contamination , at least three times, each time the container and equipment are replaced.
[0028] 2. Mechanically crush the tissue, add 4-5 times the volume of 0.01-1% collagenase digestion solution, stir at 37°C, collect the digestion solution after 4-12 hours, filter through a 200-mesh filter, add 2 times the volume of the cell suspension to wash Liquid (DMEM / F12) washed twice, using low fetal bovine serum mesenchymal medium (MesenPRO RS TM Medium, invitrogen) was resuspended and inoculated in a culture bottle, and the medium was changed in full after 24-72 hours, and half of the medium was changed every 3 days.
[0029] 3. When the cells gr...
Embodiment 2
[0038] Example 2: Preparation method of placenta-derived mesenchymal stem cells
[0039]1. Take the placenta of a healthy full-term baby and wash the surface with saline solution containing gentamicin (20,000-40,000 U / L) and heparin (50-200 U / ml) to remove residual blood and possible bacterial contamination. At least three times, each time the container and instrument are replaced.
[0040] 2. Mechanically crush the tissue, add 4-5 times the volume of collagenase digestion solution, stir at 37°C, collect the digestion solution after 4-12 hours, filter through a 200-mesh filter, add 2 times the volume of washing solution (DMEM / F12) washed twice, using low fetal bovine serum mesenchymal medium (MesenPRORS TM Medium, invitrogen) was resuspended and inoculated in a culture bottle, and the medium was changed in full after 24-72 hours, and half of the medium was changed every 3 days.
[0041] 3. When the cells grow to 80-90% confluence, carry out subculture operation. Discard t...
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