Preparation method of stem cell active factor preparation for treating endometrial injury
A technology of active factor and endometrium, applied in the field of biopharmaceuticals, can solve the problems of poor endometrial treatment effect, achieve remarkable therapeutic effect, simple culture operation, and good cell shape
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Embodiment 1
[0037] The preparation of umbilical cord mesenchymal stem cell active factor concentrate comprises the following steps:
[0038] Place the collected fresh umbilical cord in a petri dish filled with 100mL collection solution, and set aside;
[0039] Use surgical scissors to cut a 5cm umbilical cord segment, transfer the umbilical cord segment to a sterilized petri dish, add ethanol solution with a mass fraction of 75%, soak the umbilical cord in the ethanol solution and turn it over, sterilize for 45 seconds, and remove the sterilized umbilical cord The arteries and veins on the segment were washed with buffer A until there was no blood clot; after draining the water, put the processed umbilical cord segment into a flask and cut it into pieces to obtain a tissue block with a diameter of 1 mm; inoculate the tissue block into a DMEM In the culture dish of the culture medium, make the distance between each tissue block <1cm. After inoculation, place the culture dish with the tissu...
Embodiment 2
[0045] The preparation of umbilical cord mesenchymal stem cell active factor concentrate comprises the following steps:
[0046] Place the collected fresh umbilical cord in a petri dish filled with 200mL collection solution, and set aside;
[0047] Use surgical scissors to cut a 10cm umbilical cord segment, transfer the umbilical cord segment to a sterilized petri dish, add ethanol solution with a mass fraction of 75%, soak the umbilical cord in the ethanol solution and turn it over, sterilize for 60 seconds, and remove the sterilized umbilical cord The arteries and veins on the segment were washed with buffer A until there was no blood clot; after draining the water, put the processed umbilical cord segment into a flask and cut it into pieces to obtain a tissue block with a diameter of 3 mm; inoculate the tissue block into a DMEM In the culture dish of the culture medium, make the distance between each tissue block <1cm. After inoculation, place the culture dish with the tiss...
Embodiment 3
[0053] The preparation of umbilical cord blood stem cell active factor concentrate comprises the following steps:
[0054] After the placenta falls off at the end of production, insert the needle of the blood collection bag containing anticoagulant into the umbilical vein, shake the blood bag while collecting, to fully mix the blood and anticoagulant, store it at low temperature, and use it for later use;
[0055] The umbilical cord blood was separated by the density gradient method and mixed with an equal amount of normal saline to obtain a blood dilution. The blood dilution and the separation were mixed at a volume ratio of 2:1, and centrifuged for 15 minutes under the condition of a centrifugal force of 600g. The lower layer is divided into plasma layer, buffy coat layer and erythrocyte layer. Absorb the buffy coat layer and resuspend it with normal saline to obtain the first resuspension; centrifuge the first resuspension under the condition of centrifugal force of 300g for...
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