Method and key points for isolation of human amniotic epithelial cells with high yield, viability and purity

BMC Research Notes
Hossein MotedayyenAbbas Rezaei

Abstract

Human amniotic epithelial cells (hAECs) which are isolated from the amniotic membrane have stem cell-like properties and immunomodulatory effects. Several protocols have been proposed for isolation of hAECs, nevertheless, there is no report concerning isolation of highly viable hAECs, with desirable yield, and without significant purity reduction. In the current study, a detailed protocol with some modification of previous ones is presented in which the amendments led to isolation of hAECs with high purity, yield and viability. Moreover, isolated hAECs were subjected to immuno-phenotyping and their physiological status was assessed using a proliferation assay. The average yield of obtained hAECs using the new modified method was 190 × 106 cells with a mean viability of 87%, with less than 1% contamination with mesenchymal stem cells (MSCs). The isolated cells were > 95% positive for the epithelial cell markers. The lowest initial plating efficiency of the cells was 80%. Freshly isolated hAECs had the ability to proliferate for 5-6 passages in a standard culture medium.

References

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Citations

Apr 7, 2020·Journal of Biomaterials Science. Polymer Edition·Ilyas InciHuseyin Avci
Feb 2, 2021·Frontiers in Bioengineering and Biotechnology·Marline KirschAntonina Lavrentieva
Oct 17, 2020·Stem Cell Research & Therapy·Qiuwan Zhang, Dongmei Lai
Jun 10, 2021·Reproductive Sciences·Banghyun LeeHee Joong Lee

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Methods Mentioned

BETA
cesarean section
flow cytometry

Software Mentioned

CellQuest

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