Welcome to visit Acta Anatomica Sinica! Today is
Chinese
Establishment of a mouse embryonic germ cell line and preliminary study of the expression of imprinted genes
HU Jing ZHAO Qiao-shi GU Yan-li BAI Guang-yu WU Xi LEI Lei*
Acta Anatomica Sinica ›› 2013, Vol. 44 ›› Issue (3) : 397-402.
Establishment of a mouse embryonic germ cell line and preliminary study of the expression of imprinted genes
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.
Embryonic germ cell / Imprinted gene / Real-time PCR / Mouse
［1］Lawson KA, Hage WJ. Clonal analysis of the origin of primordial germ cells in the mouse ［J］. Ciba Found Symp, 1994, 182: 68-84.
［2］Durcova-Hills G, Tang F, Doody G, et al. Reprogramming primordial germ cells into pluripotent stem cells［J］. PLoS One, 2008, 3 (10):e3531.
［3］Labosky PA, Barlow DP, Hogan BL. Mouse embryonic germ (EG) cell lines: transmission through thegermline and differences in the methylation imprint of insulin-like growth factor 2 receptor (Igf2r) gene compared with embryonic stem (ES) cell lines［J］. Development, 1994, 120(11): 3197-3204.
［4］Tada T, Tada M, Hilton K, et al. Epigenotype switching ofimprintable loci in embryonic germ cells［J］. Dev Genes Evol, 1998, 207(8): 551-561.
［5］Durcova-Hills G, Burgoyne P, McLaren A.Analysis of sex differences in EGC imprinting［J］. Dev Biol, 2004, 268(1): 105-110.
［6］Hajkova P, Erhardt S, Lane N, et al. Epigenetic reprogramming in mouse primordial germ cells［J］. Mech Dev, 2002,117: 15-23.
［7］Meng GL, Teng L, Xue YF, et al. Researches on the methodology of establishment of BALB/c murine embryonic stem cell lines［J］. Acta Genetica Sinica, 2002, 29(7): 581-588. (in Chinese)
孟国良, 滕路, 薛友纺, 等. BALB/c 小鼠胚胎干细胞系建立的方法学探讨［J］.遗传学报, 2002, 29(7): 581-588.
［8］Guan N, Xu YN, Zhang QH, et al. The progress of epigenetics in somatic nuclear reprogramming［J］. Progress in Biochemistry and Biophysics, 2008, 35 (5): 1-6.(in Chinese)关娜, 徐燕宁, 张庆华, 等.体细胞核重编程中表观遗传学的研究进展［J］.生物化学与生物物理学进展, 2008, 35 (5):1-6.
［9］Buehr M. The primordial germ cells of mammals: some current perspectives［J］. Exp Cell Res, 1997, 232(2): 194-207.
［10］Denis H. A parallel between development and evolution: germ cell recruitment by the gonads［J］. Bioessays, 1994, 16(12): 933-938.
［11］MeLaren A. Germline and soma: interactions during early mouse development［J］. Semin Dev Biol, 1994, 5(1): 43-49.
［12］Shim SW, Han DW, Yang JH, et al. Derivation of embryonic germ cells from post migratory primordial germ cells, and methylation analysis of their imprinted genes by bisulfite genomic sequencing［J］. Mol Cells, 2008,25(3):358-367.
［13］Tanimoto Y, Iijima S, Hasegawa Y, et al. Embryonic stem cells derived from C57BL/6J and C57BL/6N mice［J］. Comp Med, 2008, 58(4):347-352.
［14］Lucifero D, Mertineit C, Clarke HJ, et al. Methylation dynamics of imprinted genes in mouse germ cells［J］. Genomics, 2002, 79(4): 530-538.
［15］Li C, Chen ZS, Liu Z, et al. Correlation of expression and methylation of imprinted genes with pluripotency of parthenogenetic embryonic stem cells［J］. Hum Mol Genet, 2009, 18(12): 2177-2187.
［16］Schumacher A, Doerfler W. Influence ofin vitro manipulation on the stability of methylation patterns in the Snurf/ Snrpn-imprinting region in mouse embryonic stem cells［J］.Nucleic Acids Res, 2004, 32(4): 1566-1576.
/
| 〈 |
|
〉 |
