Preparation method and application of composite decellularized amniotic membrane of endometrial stem cells
A technology of endometrial stem cells and decellularization, which is applied in the field of preparation of endometrial stem cells combined with decellularized amnion, can solve the problems of endocrine interference in patients, achieve good repair effect and avoid interference
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Embodiment 1
[0032] S1) The fresh full-term placenta was repeatedly washed with normal saline until there was no obvious blood on the surface, and then the amnion and chorion were peeled off to leave the complete amnion tissue including the epithelial cell layer, basement membrane and avascular matrix, and then the mass fraction was Rinse the intact amnion tissue once with 3% hydrogen peroxide solution, and then rinse twice with PBS solution with a pH of 7.2-7.4;
[0033] S2) Treat the intact amniotic membrane tissue treated by S1) with 0.2% EDTA at 35°C for 30 minutes, then soak it in 0.3mol / L sodium hydroxide solution for 20s, and then transfer it to 3% concentration Shake vigorously in ammonium chloride to remove cells and make translucent amniotic membrane;
[0034] S3) Wash the translucent amniotic membrane twice with PBS solution with a pH of 7.2-7.4, and then immerse and sterilize it in physiological saline containing antibiotics for 10 minutes; wherein, the antibiotics contained in...
Embodiment 2
[0040] S1) The fresh full-term placenta was repeatedly washed with normal saline until there was no obvious blood on the surface, and then the amnion and chorion were peeled off to leave the complete amnion tissue including the epithelial cell layer, basement membrane and avascular matrix, and then the mass fraction was Rinse the intact amnion tissue twice with 4% hydrogen peroxide solution, and then rinse three times with PBS solution with a pH of 7.2-7.4;
[0041] S2) Treat the intact amniotic tissue treated by S1) with 0.4% EDTA at 36°C for 40 minutes, then soak in 0.4mol / L sodium hydroxide solution for 25s, and then transfer to 4% concentration Shake vigorously in ammonium chloride to remove cells and make translucent amniotic membrane;
[0042] S3) The translucent amniotic membrane was washed three times with PBS solution with a pH of 7.2-7.4, and then soaked and disinfected in physiological saline containing antibiotics for 13 minutes; wherein, the antibiotics contained ...
Embodiment 3
[0048] S1) The fresh full-term placenta was repeatedly washed with normal saline until there was no obvious blood on the surface, and then the amnion and chorion were peeled off to leave the complete amnion tissue including the epithelial cell layer, basement membrane and avascular matrix, and then the mass fraction was Rinse the intact amnion tissue twice with 5% hydrogen peroxide solution, and then rinse four times with PBS solution with a pH of 7.2-7.4;
[0049] S2) Treat the intact amniotic tissue treated by S1) with 0.5% EDTA at 37°C for 45 minutes, then soak in 0.5mol / L sodium hydroxide solution for 30s, and then transfer to 5% concentration Shake vigorously in ammonium chloride to remove cells and make translucent amniotic membrane;
[0050] S3) The translucent amniotic membrane was washed 4 times with PBS solution with a pH of 7.2-7.4, and then soaked and disinfected in physiological saline containing antibiotics for 15 minutes; wherein, the antibiotics contained in th...
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