BMC Res Notes. 2017 Nov 2;10(1):552. doi: 10.1186/s13104-017-2880-6.

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

Motedayyen H¹, Esmaeil N¹, Tajik N², Khadem F¹, Ghotloo S³, Khani B⁴, Rezaei A⁵.

Abstract

OBJECTIVE:

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.

RESULTS:

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.