Heterozygous Embryonic Stem Cell Lines Derived from Nonhuman Primate Parthenotes

Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploi...

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Published in:	Stem cells (Dayton, Ohio) Vol. 26; no. 3; pp. 756 - 766
Main Authors:	Dighe, Vikas, Clepper, Lisa, Pedersen, Darlene, Byrne, James, Ferguson, Betsy, Gokhale, Sumita, Penedo, M. Cecilia T., Wolf, Don, Mitalipov, Shoukhrat
Format:	Journal Article
Language:	English
Published:	Bristol John Wiley & Sons, Ltd 01.03.2008
Subjects:	Animals Cell Differentiation Cell Line Cell Separation DNA Methylation Embryo, Mammalian Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Female Fertilization in Vitro Gene Expression Regulation Genomic Imprinting Genotype Heterozygote Histocompatibility Histocompatible Humans Imprinting Macaca mulatta Macaca mulatta - embryology Meiotic recombination Microsatellite Repeats - genetics Parthenogenesis Parthenogenetic Polymorphism, Single Nucleotide - genetics Prader-Willi Syndrome - genetics Primates
ISSN:	1066-5099, 1549-4918, 1549-4918
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Abstract	Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC‐derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line‐dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients. Disclosure of potential conflicts of interest is found at the end of this article.
AbstractList	Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC-derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line-dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients. Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC-derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line-dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients.Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC-derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line-dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients. Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC-derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line-dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients. Disclosure of potential conflicts of interest is found at the end of this article. Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC-derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line-dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients. Disclosure of potential conflicts of interest is found at the end of this article.
Author	Wolf, Don Ferguson, Betsy Pedersen, Darlene Mitalipov, Shoukhrat Clepper, Lisa Byrne, James Penedo, M. Cecilia T. Gokhale, Sumita Dighe, Vikas
AuthorAffiliation	a Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, Oregon, USA e Department of Obstetrics and Gynecology, Oregon Health & Science University, Beaverton, Oregon, USA b Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA c Veterinary Genetics Laboratory, University of California, Davis, California, USA d Oregon Stem Cell Center, Oregon Health & Science University, Beaverton, Oregon, USA
AuthorAffiliation_xml	– name: e Department of Obstetrics and Gynecology, Oregon Health & Science University, Beaverton, Oregon, USA – name: c Veterinary Genetics Laboratory, University of California, Davis, California, USA – name: a Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, Oregon, USA – name: b Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA – name: d Oregon Stem Cell Center, Oregon Health & Science University, Beaverton, Oregon, USA
Author_xml	– sequence: 1 givenname: Vikas surname: Dighe fullname: Dighe, Vikas – sequence: 2 givenname: Lisa surname: Clepper fullname: Clepper, Lisa – sequence: 3 givenname: Darlene surname: Pedersen fullname: Pedersen, Darlene – sequence: 4 givenname: James surname: Byrne fullname: Byrne, James – sequence: 5 givenname: Betsy surname: Ferguson fullname: Ferguson, Betsy – sequence: 6 givenname: Sumita surname: Gokhale fullname: Gokhale, Sumita – sequence: 7 givenname: M. Cecilia T. surname: Penedo fullname: Penedo, M. Cecilia T. – sequence: 8 givenname: Don surname: Wolf fullname: Wolf, Don – sequence: 9 givenname: Shoukhrat surname: Mitalipov fullname: Mitalipov, Shoukhrat email: mitalipo@ohsu.edu
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Snippet	Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for...
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SubjectTerms	Animals Cell Differentiation Cell Line Cell Separation DNA Methylation Embryo, Mammalian Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Female Fertilization in Vitro Gene Expression Regulation Genomic Imprinting Genotype Heterozygote Histocompatibility Histocompatible Humans Imprinting Macaca mulatta Macaca mulatta - embryology Meiotic recombination Microsatellite Repeats - genetics Parthenogenesis Parthenogenetic Polymorphism, Single Nucleotide - genetics Prader-Willi Syndrome - genetics Primates
Title	Heterozygous Embryonic Stem Cell Lines Derived from Nonhuman Primate Parthenotes
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