The human AEC isolation was performed as previously described.30 –32 (no link found, link) Briefly, the amnion membrane was removed from the inner layer of the placenta from full-term healthy pregnancies, and washed extensively to remove blood. The amnion membrane was immersed in TrypLE 10x (Gibco) for 30 min at 37°C to release mainly AEC from the amnion membrane. The released AEC were collected and washed by serial centrifugation and filtered through a 100 µm cell strainer to remove undigested tissues or large clamps. Cell viability and yield were determined by the Trypan Blue exclusion and cells immediately exposed to cryopreservation procedure.
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Corresponding Organization : King's College London
Other organizations :
Karolinska Institutet, London Centre for Nanotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Karolinska University Hospital
Removal of amnion membrane from the inner layer of the placenta
Washing of the amnion membrane to remove blood
Immersion of the amnion membrane in TrypLE 10x for 30 min at 37°C
Filtration of the released AEC through a 100 µm cell strainer
dependent variables
AEC yield
AEC viability
control variables
Full-term healthy pregnancies
Cryopreservation procedure
controls
No positive or negative controls were explicitly mentioned.
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