Construction method of medical human amniotic membrane tissue reserve bank
A construction method and human amniotic membrane technology are applied in the field of construction of human tissue reserve bank, which can solve the problem of amniotic membrane resource loss and other problems, and achieve the effects of low construction cost, rich application prospects, and simple and easy operation.
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Embodiment 1
[0028] The construction method of the medical human amniotic membrane tissue reserve of the present invention comprises the following steps:
[0029] (1) The selection of human amniotic membrane strictly follows the medical standards of donors and establishes data files;
[0030] (2) Take the fetal membranes of the donor who meet the medical standards of caesarean section, and bluntly separate the amniotic membranes from the fetal membranes; if figure 1 shown.
[0031] (3) After repeated rinsing with 0.9% saline, cut into a rectangular membrane of a certain size, such as 10×12cm 2 And mark the surface layer of basement membrane with sterile suture;
[0032] (4) Soak in 0.9% normal saline containing 1000 U / ml gentamicin and 2.5 μg / ml amphotericin B for 30 minutes;
[0033] (5) Put it into a 75ml sterile cell culture bottle containing 4% glycerol and 0.9% normal saline mixture, and store it in a deep-low temperature refrigerator at -80°C;
[0034] (6) The human amniotic mem...
Embodiment 2
[0039] (1) The selection of human amniotic membrane strictly follows the medical standards of donors and establishes data files;
[0040] (2) Take the fetal membranes of the donor that meet the medical standards and give birth normally, and bluntly separate the amniotic membranes from the fetal membranes;
[0041] (3) After repeated rinsing with phosphate buffered saline (PBS) water, cut a circle with a diameter of 7 cm and mark the surface layer of the basement membrane with sterile sutures;
[0042] (4) Soak in phosphate buffered saline (PBS) containing 1000 U / ml gentamicin and 2.5 μg / ml amphotericin B for 40 minutes;
[0043] (5) Put it into a 150ml plastic bag containing 10% glycerol-phosphate buffered saline (PBS) mixture, and store it in a deep-low temperature refrigerator at -80°C;
[0044] (6) The human amniotic membrane tissue membrane prepared by the above method is sterilized by cobalt 60 irradiation, and the irradiation dose is 25kGy to obtain a medical human amni...
Embodiment 3
[0047] (1) The selection of human amniotic membrane strictly follows the medical standards of donors and establishes data files;
[0048] (2) Take the fetal membranes of the donor who meet the medical standards of caesarean section, and bluntly separate the amniotic membranes from the fetal membranes;
[0049] (3) After repeated rinsing with 0.9% normal saline, cut to a diameter of 19×19cm 2 And use sterile sutures to mark the basement membrane surface layer;
[0050] (4) Soak in 0.9% normal saline containing 1000 U / ml gentamicin and 2.5 μg / ml amphotericin B for 30 minutes;
[0051] (5) Put it into a 250ml sterile cell culture bottle filled with 0.9% normal saline solution containing 100% glycerol that is ready to be sterilized, and store it in a deep-low temperature refrigerator at -80°C;
[0052] (6) The human amniotic membrane tissue membrane prepared by the above method is sterilized by cobalt 60 irradiation, and the irradiation dose is 25kGy to obtain a medical human ...
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