Method for preparing mesenchymal stem cell
A technique for stromal stem cells and cell culture, applied in the field of mesenchymal stem cell preparation, which can solve the problems of reduced stem cell number, proliferation and differentiation ability, inability to extract bone marrow, and patient injury.
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Embodiment 1
[0037] Embodiment 1 (tissue block adherence method):
[0038] (1) Umbilical cord pretreatment
[0039] Take the fresh umbilical cord of the newborn, ligate both ends of the umbilical cord, put it into 100ml of RPMI-1640 culture solution containing 100U / ml penicillin and 100mg / ml streptomycin, and transport it at 4°C; Soak the umbilical cord in % medical alcohol for 45 seconds, remove about 2 cm from both ends of the ligated umbilical cord, cut the middle section into 1.5 cm long pieces, and obtain 20 pieces in total. Rinse the small section of umbilical cord repeatedly with normal saline to remove residual blood clots, cut open the small section of umbilical cord, remove the umbilical arterial vein, rinse repeatedly with normal saline until there is no residual blood, and cut into pieces to 1mm 3 , get a small piece of umbilical cord tissue with a volume of about 30ml, and refrigerate at 4°C;
[0040] (2) Cryopreservation of umbilical cord tissue blocks
[0041] Prepare 30m...
Embodiment 2
[0054] Embodiment 2 (enzyme digestion method):
[0055] (1) Umbilical cord pretreatment
[0056] Take the fresh umbilical cord of the newborn, ligate both ends of the umbilical cord, put it into 100ml of RPMI-1640 culture solution containing 100U / ml penicillin and 100mg / ml streptomycin, and transport it at 4°C; Soak the umbilical cord in % medical alcohol for 45 seconds, remove about 2 cm from both ends of the ligated umbilical cord, cut the middle section into 1.5 cm long pieces, and obtain 20 pieces in total. Rinse the small section of umbilical cord repeatedly with normal saline to remove residual blood clots, cut open the small section of umbilical cord, remove the umbilical arterial vein, rinse repeatedly with normal saline until there is no residual blood, and cut into pieces to 1mm 3 , get a small piece of umbilical cord tissue with a volume of about 30ml, and refrigerate at 4°C;
[0057] (2) Cryopreservation of umbilical cord tissue blocks
[0058] Take out 30ml of ...
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