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Efficient production of androgenetic embryos by round spermatid injection
Author(s) -
Miki Hiromi,
Hirose Michiko,
Ogonuki Narumi,
Inoue Kimiko,
Kezuka Fuyuko,
Honda Arata,
Mekada Kazuyuki,
Hanaki KenIchi,
Iwafune Hirotaka,
Yoshiki Atsushi,
Ishino Fumitoshi,
Ogura Atsuo
Publication year - 2009
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20451
Subject(s) - spermatid , biology , embryo , andrology , enucleation , oocyte , human fertilization , fetus , fertilisation , embryogenesis , genetics , reproductive technology , pregnancy , sperm , medicine
Mammalian androgenetic embryos can be produced by pronuclear exchange of fertilized oocytes or by dispermic in vitro fertilization of enucleated oocytes. Here, we report a new technique for producing mouse androgenetic embryos by injection of two round spermatid nuclei into oocytes, followed by female chromosome removal. We found that injection of round spermatids resulted in high rates of oocyte survival (88%). Androgenetic embryos thus produced developed into mid‐gestation fetuses at various rates, depending on the mouse strain used. All the fetuses examined maintained paternally specific genomic imprinting memories. This technique also enabled us to produce complete heterozygous F1 embryos by injecting two spermatids from different strains. The best rate of fetal survival (12% per embryos transferred) was obtained with C57BL/6 × DBA/2 androgenetic embryos. We also generated embryonic stem cell lines efficiently with the genotype of Mus musculus domesticus × M. m. molossinus . Thus, injection of two round spermatid nuclei followed by maternal enucleation is an effective alternative method of producing androgenetic embryos that consistently develop into blastocysts and mid‐gestation fetuses. genesis 47:155–160, 2009. © 2009 Wiley‐Liss, Inc.