Comparison of Germ Cell Gene Expressions in Spontaneous Monolayer versus Embryoid Body Differentiation of Mouse Embryonic Stem Cells toward Germ Cells

• Published in: International journal of fertility & sterility Vol. 13; no. 2; pp. 139 - 147
• Main Authors: Gholamitabar Tabari, Maryam, Jorsaraei, Seyed Gholam Ali, Ghasemzadeh-Hasankolaei, Mohammad, Ahmadi, Ali Asghar, Ghasemi, Masoumeh
• Format: Journal Article
• Language: English
• Published: Iran Royan Institute of Iran 01-07-2019; Royan Institute; Royan Institute (ACECR), Tehran
• Subjects: Amino acids; Cell cycle; differentiation; embryoid body; embryonic stem cells; Embryos; Gene expression; Genes; Leukemia; Medical research; monolayer; Morphology; Original; Polymerase chain reaction; Stem cells; United Kingdom > UK; United States > US; Japan; Germany; Monolayer; Embryonic Stem Cells; Differentiation; Embryoid Body
• ISSN: 2008-076X; 2008-0778
• DOI: 10.22074/ijfs.2019.5557

Genetic and morphologic similarities between mouse embryonic stem cells (ESCs) and primordial germ cells (PGCs) make it difficult to distinguish differentiation of these two cell types
. Using specific GC markers expressed in low level or even not expressed in ESCs- can help recognize differentiated cells
. We attempted to differentiate the mouse ESCs into GC-like cells spontaneously in monolayer and EB culture method.
In this experimental study, we attempted to differentiate ESCs, Oct4-GFP OG2, into GC-like cells (GCLCs) spontaneously in two different ways, including: i. Spontaneous differentiation of ESCs in monolayer culture as (SP) and ii. Spontaneous differentiation of ESCs using embryoid body (EB) culture method as (EB+SP). During culture, expression level of four GC specific genes
and
were evaluated.
In both groups,
was down-regulated (P=0.3), while
and
were up-regulated (P=0.3 and P=0.04, respectively).
and
were decreased in EB+SP and they were increased in SP group, while no significant difference was determined between them (P=0.1, P=0.07). Additionally, in SP group, gene expression of
and
were up-regulated and they had significant differences compared to EB+SP group (P=0.00 and P=0.01, respectively).
was down-regulated in the both groups. Flow-cytometry analysis showed that mean number of Mvh-positive cells in the SP group was significantly greater compared to ESCs, EB+SP and EB7 groups (P=0.00, P=0.01, and P=0.3, respectively).
These findings showed that ESCs were differentiated into GCLCs in both group. But spontaneous differentiation of ESCs into GCLCs in SP group (monolayer culture) compared to EB+SP (EB culture methods) has more ability to express GCs markers.