Method for removing cells from human amnion
A decellularization and human amniotic membrane technology, applied in artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of low decellularization efficiency, inability to completely retain the extracellular matrix of amniotic membrane, and narrow application range.
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Embodiment 1
[0014] Embodiment 1: A method for decellularization of human amnion, including the following specific processes:
[0015] (1) Remove blood clots and chorion from the fresh amniotic membrane of human placenta, leaving the complete amniotic membrane tissue including epithelial cell layer, basement membrane and avascular matrix, and wash it with phosphate buffer solution (PBS) with a pH value of 7.4, Obtain a clean translucent amniotic membrane;
[0016] (2) Rinse the translucent amniotic membrane obtained above twice with alcohol with a volume concentration of 75%, and then rinse twice with PBS, then mix penicillin and streptomycin into PBS to form a double antibody / PBS solution, in which penicillin The content of streptomycin and streptomycin in PBS is 100U / L, and then the translucent amniotic membrane is placed in the double antibody / PBS solution with a volume ratio of 1:1, soaked and shaken, and the medium is changed every 12 hours, twice in total ;
[0017] (3) Mix the dou...
Embodiment 2
[0020] Embodiment 2: A method for decellularization of human amnion, including the following specific processes:
[0021] (1) Remove blood clots and chorion from the fresh amniotic membrane of human placenta, leaving the complete amniotic membrane tissue including epithelial cell layer, basement membrane and avascular matrix, and wash it with phosphate buffer solution (PBS) with a pH value of 7.4, Obtain a clean translucent amniotic membrane;
[0022] (2) Rinse the translucent amniotic membrane obtained above once with 75% alcohol by volume, and rinse four times with PBS, then mix penicillin and streptomycin into PBS to form a double antibody / PBS solution, in which penicillin The contents of streptomycin and streptomycin in PBS are both 200U / L, and then the translucent amniotic membrane is placed in the double antibody / PBS solution with a volume ratio of 1:3, soaked and shaken, and the medium is changed every 12 hours, twice in total ;
[0023] (3) Mix the double antibody an...
Embodiment 3
[0026] Embodiment 3: A method for decellularization of human amniotic membrane, including the following specific processes:
[0027] (1) Remove blood clots and chorion from the fresh amniotic membrane of human placenta, leaving the complete amniotic membrane tissue including epithelial cell layer, basement membrane and avascular matrix, and wash it with phosphate buffer solution (PBS) with a pH value of 7.4, Obtain a clean translucent amniotic membrane;
[0028] (2) Rinse the translucent amniotic membrane obtained above twice with alcohol with a volume concentration of 75%, and then rinse three times with PBS, and then mix penicillin and streptomycin into PBS to form a double antibody / PBS solution, in which penicillin The content of streptomycin and streptomycin in PBS is 300U / L, and then the translucent amniotic membrane is placed in the double antibody / PBS solution with a volume ratio of 1:2, soaked and shaken, and the medium is changed every 12 hours, twice in total ;
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