Comparison of Germ Cell Gene Expressions in Spontaneous Monolayer versus Embryoid Body Differentiation of Mouse Embryonic Stem Cells toward Germ Cells
Authors
Abstract:
Objective Genetic and morphologic similarities between mouse embryonic stem cell (ESCs) and Primordial Germ Cell (PGCs) make it difficult to distinguish the two cell types in in vitro differentiation. Using the expression of specific markers of germ cells that are not expressed or expressed at low levels in ESCs, can help recognizing in vitro differentiated cells MaterialsAndMethods In this study, we attempted to differentiate the ESCs, Oct4-GFP OG2 into Germ Cell-like cell (GCLCs) spontaneously in two different ways:1) Spontaneous differentiation of ESCs in monolayer culture as (SP), and 2) Spontaneous differentiation of ESCs using embryoid body (EB) culture method as (EB+SP). During culture duration, the expression level of four germ cell specific genes (Fkbp6, Mov10l1, Riken, Tex13) and Mvh, Scp3, Stra8, Oct4 were evaluated. Results In both groups, Mov10l1 was down regulated (p=0.3) and Tex13 and Riken were up regulated (p=0.3, p=0.04), respectively. Fkbp6 and Stra8 were decreased in EB+SP and increased in SP group with no significant differences between them (p=0.1, p=0.07). Additionally, in SP group, gene expression of Mvh and scp3 were up regulated and had significant differences compared with EB+SP group (p=0.00, p= 0.01), respectively. Oct4 was down regulated in both groups. Flow cytometry analysis showed that mean number of Mvh-positive cells in the SP group was greater significantly as compared with ESCs, EB+SP and EB7 groups (p=0.00, p=0.01, p=0.3, respectively). Conclusion The findings showed that ESCs differentiated in to GCLCs in both group. But spontaneous differentiation of ESCs into GCLCs in SP group (monolayer culture) compared with EB+SP (EB culture methods) has more ability to expression of germ cells markers.
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Journal title
volume 13 issue 2
pages 139- 147
publication date 2019-07-01
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