Establishment of a mouse embryonic germ cell line and preliminary study of the expression of imprinted genes

doi:10.3969/j.issn.0529-1356.2013.03.021

Abstract

Abstract:

Objective To establish a mouse embryonic germ cell (EGCs) line and to detect the expression of imprinted genes
in order to provide basic information for further study and application of embryonic germ cells. Methods EGCs were isolate
from primordial germ cells collected from the genital ridge of 12.5 days postcoitum (dpc). The pluripotent characteristics of
the established EGCs were detected by alkaline phophatase (AKP) staining, immunofluorescent detection of mouse embryonic stem
cells (ESCs) surface antigens, and cell differentiations in vivo. The expressions of several patrilineal and matrilineal
imprinted genes, such as Ins2, Lgf2, H19, Lgf2r and so forth, were also detected by quantitative reverse transcriptase-polymerase
chain reaction in both EGCs and ESCs. Results The EGCs showed positive for alkaline phosphatase. The pluripotency marker Oct4
and the cell surface marker SSEA-1 were also shown in EGCs cells. Karyotype analysis indicted that EGCs had normal 40 chromosomes,
and differentiated into the tissues presenting three germinal layers derivations in vivo, suggesting that embryonic germ cells
had pluripotent characteristics. Real-time PCR showed that the expression levels of imprinted genes in EGCs were significantly
highter compared with those in ESCs. Conclusion The genomic imprinting memories in EGCs generated from primordial germ cells
which collected from the genital ridge of 12.5 dpc are completely erased.

Key words: Embryonic germ cell, Imprinted gene, Real-time PCR, Mouse

CLC Number:

R329.2

HU Jing ZHAO Qiao-shi GU Yan-li BAI Guang-yu WU Xi LEI Lei*. Establishment of a mouse embryonic germ cell line and preliminary study of the expression of imprinted genes[J]. AAS, 2013, 44(3 ): 397-402.

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