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a Department of Obstetrics & Gynecology, Faculty of Medicine, University of Tokyo, Tokyo, Japan;
b Division of Reproductive Sciences, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, Oregon, USA;
c Stem Cell Program, Institute of Zoology, Genomics Research Center, Academia Sinica, Nankang, Taipei, Taiwan
Key Words. Imprinting • Embryonic stem cells • Rhesus monkey • Embryos
Correspondence: Don P. Wolf, Ph.D., Division of Reproductive Sciences, Oregon National Primate Research Center, 505 N.W. 185th Ave., Beaverton, Oregon 97006, USA. Telephone: 503-690-5326; Fax: 503-533-2494; e-mail: wolfd{at}ohsu.edu
Received July 6, 2005;
accepted for publication October 23, 2005.
Genomic imprinting involves modification of a gene or a chromosomal region that results in the differential expression of parental alleles. Disruption or inappropriate expression of imprinted genes is associated with several clinically significant syndromes and tumorigenesis in humans. Additionally, abnormal imprinting occurs in mouse embryonic stem cells (ESCs) and in clonally derived animals. Imprinted gene expression patterns in primate ESCs are largely unknown, despite the clinical potential of the latter in the cell-based treatment of human disease. Because of the possible implications of abnormal gene expression to cell or tissue replacement therapies involving ESCs, we examined allele specific expression of four imprinted genes in the rhesus macaque. Genomic and complementary DNA from embryos and ESC lines containing useful single nucleotide polymorphisms were subjected to polymerase chain reaction–based amplification and sequence analysis. In blastocysts, NDN expression was variable indicating abnormal or incomplete imprinting whereas IGF2 and SNRPN were expressed exclusively from the paternal allele and H19 from the maternal allele as expected. In ESCs, both NDN and SNRPN were expressed from the paternal allele while IGF2 and H19 showed loss of imprinting and biallelic expression. In differentiated ESC progeny, these expression patterns were maintained. The implications of aberrant imprinted gene expression to ESC differentiation in vitro and on ESC-derived cell function in vivo after transplantation are unknown.
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