Generation and characterization of mouse parthenogenetic embryonic stem cells containing genomes from non‐growing and fully grown oocytes

Abstract

It is known that oocytes can be activated without male contribution in vitro and develop to blastocysts which are used to isolate parthenogenetic embryonic stem cells. Unfortunately, differentiation capacity of the parthenogenetic embryonic stem cells was rather lower than fertilized embryos derived ES cells, which might be the result of the absence of male genome. It had been found that some maternally expressed genes were repressed and some paternally expressed genes were expressed in the non-growing oocytes. Therefore, maternal genome from non-growing oocytes can partially act as "sperm genome". In the present study, parthenogenetic blastocysts containing genome from non-growing and fully grown oocytes (named as NF-pBlastocysts) were produced by germinal vesicle transfer, and three newly established parthenogenetic embryonic stem (named as NF-pES) cell lines were derived from the resulting parthenogenetic blastocysts. All three NF-pES cell lines were positive for ES cell markers, such as alkaline phosphatase (AKP), stage-specific embryonic antigen 1 (SSEA-1) and octamer-binding transcription factor (Oct-4). They have a normal chromosome karyotype (40) and can be maintained in an undifferentiated state for extended periods of time. When NF-pES cells were injected into severe combined immunodeficient mice, teratomas with all three embryonic germ layers were obtained. The in vitro differentiation potential of NF-pES cells was analyzed by embryonic bodies (EB) formation. The expression of germ layer markers, such as nestin (ectoderm), desmin (mesoderm), and alpha-fetoprotein (endoderm) demonstrated that the NF-pES cells can differentiate into all three germ layers.

Aggregation of pre‐implantation embryos improves establishment of parthenogenetic stem cells and expression of imprinted genes

“…; Shao et al . ). In addition, parthenogenetic stem cells are deficient in the expression of key paternal alleles, which could be used as a model for studying the genomic imprinting (Tada et al .…”

“…To date, PgES cells have been successfully established in many species, including mouse, rabbit, nonhuman primates, even in humans (Kaufman et al 1983;Cibelli 2002;Brevini et al 2008). It has been proved that PgES cells are similar to ES cells from fertilization (fES cells) in most aspects, including the expression of certain stem cell markers, chromatin methylation patterns, embryoid body formation, and teratoma development (Park et al 1998;Barton et al 2001;Hwang et al 2004;Shao et al 2007). In addition, parthenogenetic stem cells are deficient in the expression of key paternal alleles, which could be used as a model for studying the genomic imprinting Deng et al 2007;Horii et al 2008).…”

Chromosomal Abnormalities in Embryonic and Somatic Stem Cells

“…Chromosome deletions and translocations have also been reported in the hPES-2 cell line, when cultivated for 120 passages [Mai et al, 2007]. Few cytogenetic data are available on mouse pESC lines, reporting a stable karyotype during culture (up to passage 120), analyzed by standard G-banding [Shao et al, 2007;Ju et al, 2008;Shan et al, 2012].…”

Quantitative proteomics analysis of parthenogenetically induced pluripotent stem cells

“…Nestin is a marker for the ectoderm and plays a role in nerve development (Zimmerman et al, 1994); this protein has also been identified in mouse pES cells generated from growing oocytes (Shao et al, 2007). BPY2-interacting protein 1 interacts with ubiquitin protein ligase E3A (Wong et al, 2002) and may be involved in male germ cell development and male fertility.…”