Method for extracting and culturing amniotic stem cells
A stem cell and amniotic membrane technology, applied in the field of culture and extraction of amniotic membrane stem cells, can solve the problems of low efficiency and low purity of stem cells, and achieve the effect of high extraction efficiency and high purity
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Embodiment 1
[0027] 1. Amniotic membrane preparation
[0028] Take normal cesarean section placenta without hepatitis, syphilis, HIV and other infectious diseases.
[0029] The amniotic membrane is mechanically separated from the placenta, and then the removed amniotic membrane is placed in a PBS container containing double antibodies and washed repeatedly to remove blood and debris.
[0030] 2. Extraction of amniotic membrane stem cells
[0031] 2.1. Shake the amniotic membrane with mixed solution 1 at room temperature and soak for 0.5-1 hour, then wash it with tris buffer solution; mixed solution 1 contains 0.03% (w / v) sodium lauryl sulfate and 0.1 %(w / v)
[0032] EDTA in Tris buffer;
[0033] 2.2. Gently scrape the surface of the amniotic membrane with a cell scraper, rinse with PBS for 3 to 5 times, and mechanically cut the amniotic membrane to a size of 2*2mm;
[0034] 2.3. Add the amniotic membrane fragments to the mixture 2, and then put it in 37℃, CO 2 The volume fraction of t...
Embodiment 2
[0042] Embodiment 2 is basically the same as Embodiment 1, the difference is:
[0043] Mixed solution 1 is tris buffer solution containing 0.05% (w / v) sodium lauryl sulfate and 0.5% (w / v) ethylenediaminetetraacetic acid;
[0044] The mixed solution 2 is a mixed solution of α-MEM medium containing 3 g / L type II collagenase, 0.1 g / L DNase I and 15% fetal bovine serum.
Embodiment 3
[0046] Embodiment 2 is basically the same as Embodiment 1, the difference is:
[0047] Mixed solution 1 is tris buffer solution containing 0.04% (w / v) sodium lauryl sulfate and 0.3% (w / v) ethylenediaminetetraacetic acid;
[0048] The mixed solution 2 is a mixed solution of α-MEM medium containing 2.5g / L type II collagenase, 0.08g / L DNase I and 15% fetal bovine serum.
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