Construction method of medical human amniotic membrane tissue reserve bank

Abstract

The invention relates to a construction method of a medical human amniotic membrane tissue reserve bank, comprising the following steps of: 1, selecting materials of human amniotic membranes and establishing a file; 2, taking a fetal membrane with a donor meeting medical standard cesarean section or normal labor; separating camisia foetus and maintaining amniotic membranes; 3, repeatedly rinsing the amniotic membranes by using a phosphate buffering solution and cutting into membrane sheets; 4, utilizing a phosphate buffering solution containing gentamicin with the concentration of 1000 U / ml and amphotericin B with the concentration of 2.5 micrograms / millimeter; 5, placing the membrane sheets into a sterile cell culturing bottle or plastic bag of a phosphate buffering solution containing 4-100% of glycerol and freezing and storing at a temperature in a range of 20 DEG C below zero to 80 DEG C; 6, carrying out radiation sterilization treatment on human amniotic membrane sheets by cobalt 60 to obtain medical human amniotic membrane sheets; and 7, placing amniotic membrane tissue membrane sheets to be frozen and stored at 80 DEG C below zero to establish an amniotic membrane tissue membrane sheet file capable of being searched, namely establishing the human amniotic membrane tissue membrane sheet reserve bank which has long storage time and can be effectively used for clinical treatment in time.

Description

technical field

[0001] The invention relates to a method for constructing a human tissue reserve, in particular to a method for constructing a medical human amniotic tissue reserve. Background technique

[0002] With the continuous development of society, the continuous improvement of people's living standards and the aging of the population, chronic and difficult-to-heal wounds caused by various diseases are also increasing, which has become a serious threat to people's physical and mental health, reducing their quality of life and work, and increasing An important reason for social medical security burden. Especially for perianal diseases, not only the postoperative wound is difficult to heal, but also the patient’s wound is unbearably painful when changing the dressing, the wound is prone to bleeding or oozing, difficult to bandage, easy to be contaminated by feces, and restricts the patient’s activity for a long time, etc., causing great harm to the patient. Physical a...

Examples

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 ...

technical field

[0001] The invention relates to a method for constructing a human tissue reserve, in particular to a method for constructing a medical human amniotic tissue reserve. Background technique

[0002] With the continuous development of society, the continuous improvement of people's living standards and the aging of the population, chronic and difficult-to-heal wounds caused by various diseases are also increasing, which has become a serious threat to people's physical and mental health, reducing their quality of life and work, and increasing An important reason for social medical security burden. Especially for perianal diseases, not only the postoperative wound is difficult to heal, but also the patient’s wound is unbearably painful when changing the dressing, the wound is prone to bleeding or oozing, difficult to bandage, easy to be contaminated by feces, and restricts the patient’s activity for a long time, etc., causing great harm to the patient. Physical a...

Examples

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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