Abstract. Mouse bi-maternal embryos (BMEs) that contain two haploid sets of genomes from non-growing (ng) and fully-grown (fg) oocytes develop to embryonic day (E) 13.5. However, the ng/fg BMEs never develop beyond E13.5 because of repression of the paternally expressed imprinted genes, Igf2 and Dlk1. The present study was conducted to address the issue of whether fetal hematopoietic disorder is involved in the restricted development of BMEs. FACS analysis revealed that the livers of ng wt /fg BMEs contained increased numbers of immature c-kit + /ter119 -hematopoietic cells, were while the numbers of mature c-kit -/ter119 + hematopoietic cells were decreased. This finding was supported by histological observations. Quantitative gene expression analysis revealed that Igf2 and Dlk1 expression was repressed in the liver. To understand the role of paternally-methylated imprinted genes on chromosomes 7 and 12, particularly Igf2 and Dlk1, in fetal liver hematopoiesis, we constructed ng Δch7 /fg, ng [177][178][179][180][181][182] 2008) ue to imprinted genes, which exhibit monoallelic expression depending upon parental origin-dependent epigenetic modifications, such as DNA methylation imposed during gametogenesis, mouse parthenogenetic embryos never develop beyond day 10 of gestation in mice [1][2][3][4]. However, reconstructed bi-maternal embryos (ng/fg BMEs), which contain 2 haploid sets of genomes obtained from a non-growing stage oocyte (ng) and a fully grown oocyte (fg), develop to embryonic day (E) 13.5 [5]. A detailed gene expression analysis revealed that this extended development of ng/fg BMEs results from the appropriate expression of a majority of the imprinted genes: however, the paternally-methylated and paternally-expressed genes are still repressed [6,7]. To induce expression of Igf2 and Dlk1 from the ng oocyte allele, we reconstructed ng/fg BMEs using ng oocytes harboring a deletion in the H19 transcription unit and its differentially methylated region (Δch7) or a deletion in the methylated region IG-DMR (Δch12) in the ng allele. We demonstrated that both genotypes of ng Δch7 /fg BMEs [7] and ng Δch12 /fg BMEs [8] developed to E18.5, but also that they experienced severe growth retardation and died soon after recovery from the uterus. Furthermore, we recently demonstrated that ng/fg BMEs harboring both Δch7 and Δch12 (ΔDouble) expressed Igf2 and Dlk1 at appropriate levels and developed to normal adults at high frequency [9]. From these results, it has been suggested that both Igf2 and Dlk1 are critical for normal mouse embryonic development, while these paternally expressed imprinted genes function as a strict barrier to development beyond mid-gestation (E13.5) in mice [7,8,10]. However, phenotypical analyses for ng/fg bi-maternal fetuses have rarely been reported. In order to gain a further understanding of the features of the bi-maternal fetuses, we focused on hematopoiesis because the fetal hematopoietic system in the liver is critically important for survival and growth of mouse embryos. In mouse...