Reproductive Biology, Volume (20), No (3), Year (2020-9) , Pages (338-347)

Title : ( Investigating the expression of pluripotency-related genes in human amniotic fluid cells: A semi-quantitative comparison between different subpopulations, from primary to cultured amniocytes )

Authors: Seyed Mehdi Hoseini , Fateme Montazeri , Ahmad Reza Bahrami , Seyed Mehdi Kalantar , Saeede Rahmani , Fateme Zarein , Maryam Moghaddam Matin ,

Citation: BibTeX | EndNote

Abstract

Various classifications have been proposed for human amniotic subpopulations, including classification of spindle-shaped (SS) and round-shaped (RS) cells, as well as the more referred triple-category of pithelioid (Etype) cells, amniotic fluid-specific (AF-type) cells and fibroblastoid (F-type) cells. The present study aims to investigate these amniotic subpopulations regarding the expression of some stem cell markers, including OCT4, NANOG, SOX2, C-KIT (CD117), C-MYC, KLF4, and THY1 (CD90). Flow cytometry was performed to characterize the isolated clonal subpopulations for a hematopoietic and a mesenchymal marker using PE-CD31 and FITCCD90, respectively. A semi-quantitative RT-PCR analysis was carried out on the isolates in the second half of their lifespan when the cells were at the stationary phase of the growth curve. Characterization of isolated cells demonstrated that all clones including both epithelioid and fibroblastoid cells, had mesenchymal, not hematopoietic, lineage. RT-PCR analysis also revealed a higher expression of the target genes in epithelioid cells. Furthermore, the expression pattern of the genes and their correlations were remarkably different between primary- and long term-cultured amniocytes. Taken together, our results showed that the primary-cultured cells express the stemness genes equally, whereas the long term-cultured amniocytes exhibited a highly variable manner in the expression pattern of the genes. Diverse derivation site of amniocytes and individual genetic background can potentially explain the observed variation in the expression level of the target genes. These can also explain why amniocytes differ in many respects observed in our study, including survival rate, plastic adhesion, and growth characteristics.

Keywords

, Mesenchymal stem cells, Human amniocytes, Amniotic subpopulations, Pluripotency markers
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@article{paperid:1081480,
author = {Hoseini, Seyed Mehdi and Fateme Montazeri and Bahrami, Ahmad Reza and Seyed Mehdi Kalantar and Saeede Rahmani and Fateme Zarein and Moghaddam Matin, Maryam},
title = {Investigating the expression of pluripotency-related genes in human amniotic fluid cells: A semi-quantitative comparison between different subpopulations, from primary to cultured amniocytes},
journal = {Reproductive Biology},
year = {2020},
volume = {20},
number = {3},
month = {September},
issn = {1642-431X},
pages = {338--347},
numpages = {9},
keywords = {Mesenchymal stem cells; Human amniocytes; Amniotic subpopulations; Pluripotency markers},
}

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%0 Journal Article
%T Investigating the expression of pluripotency-related genes in human amniotic fluid cells: A semi-quantitative comparison between different subpopulations, from primary to cultured amniocytes
%A Hoseini, Seyed Mehdi
%A Fateme Montazeri
%A Bahrami, Ahmad Reza
%A Seyed Mehdi Kalantar
%A Saeede Rahmani
%A Fateme Zarein
%A Moghaddam Matin, Maryam
%J Reproductive Biology
%@ 1642-431X
%D 2020

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