Donor cell differentiation, reprogramming, and cloning efficiency: Elusive or illusive correlation?

Oback, Björn; Wells, David N.

doi:10.1002/mrd.20654

Cited by 71 publications

(53 citation statements)

References 74 publications

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“…More recently, iPSC have been generated from monkey (42), rat (43), and, while this manuscript was under review, miniature swine (44). This technology might be especially valuable in agriculturally important species for nuclear transfer experiments, as pluripotent cells might provide higher cloning efficiency and avoid the abnormalities and deaths associated with using differentiated somatic cells (45). In addition, it might be possible to establish gene targeting technologies with the ultimate goal of creating genetic models for human diseases in species where mouse models are inappropriate.…”

Section: Discussionmentioning

confidence: 99%

Derivation of induced pluripotent stem cells from pig somatic cells

Ezashi¹,

Telugu²,

Alexenko³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

439

399

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For reasons that are unclear the production of embryonic stem cells from ungulates has proved elusive. Here, we describe induced pluripotent stem cells (iPSC) derived from porcine fetal fibroblasts by lentiviral transduction of 4 human (h) genes, hOCT4, hSOX2, hKLF4, and hc-MYC, the combination commonly used to create iPSC in mouse and human. Cells were cultured on irradiated mouse embryonic fibroblasts (MEF) and in medium supplemented with knockout serum replacement and FGF2. Compact colonies of alkaline phosphatase-positive cells emerged after Ϸ22 days, providing an overall reprogramming efficiency of Ϸ0.1%. The cells expressed porcine OCT4, NANOG, and SOX2 and had high telomerase activity, but also continued to express the 4 human transgenes. Unlike human ESC, the porcine iPSC (piPSC) were positive for SSEA-1, but negative for SSEA-3 and -4. Transcriptional profiling on Affymetrix (porcine) microarrays and real time RT-PCR supported the conclusion that reprogramming to pluripotency was complete. One cell line, ID6, had a normal karyotype, a cell doubling time of Ϸ17 h, and has been maintained through >220 doublings. The ID6 line formed embryoid bodies, expressing genes representing all 3 germ layers when cultured under differentiating conditions, and teratomas containing tissues of ectoderm, mesoderm, and endoderm origin in nude mice. We conclude that porcine somatic cells can be reprogrammed to form piPSC. Such cell lines derived from individual animals could provide a means for testing the safety and efficacy of stem cell-derived tissue grafts when returned to the same pigs at a later age.iPS ͉ reprogramming ͉ OCT4

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Section: Discussionmentioning

confidence: 99%

Derivation of induced pluripotent stem cells from pig somatic cells

Ezashi¹,

Telugu²,

Alexenko³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

439

399

View full text Add to dashboard Cite

show abstract

“…We have learnt from mouse experiments that the easiest cells to reprogram are blastomeres from a morula; however, efficiency progressively declines when cells from the inner cell mass and tissue-specific cells are used (Gurdon and Melton, 2008;Hochedlinger and Jaenisch, 2006;Thuan et al, 2010;Wakamatsu, 2008). Side-by-side comparisons of different cell types showed that ESCs were more amenable to cloning than somatic fibroblasts (Rideout et al, 2000); and even among different somatic cell populations, the variations in cloning efficiency were significant (Oback, 2009;Oback and Wells, 2007). In amphibians, adult frogs were obtained when donor nuclei were isolated from blas-tomeres (Gurdon et al, 1958) and embryonic-intestinal cells (Gurdon and Uehlinger, 1966), but only tadpoles were produced using adult cells as donor nuclei (Gurdon et al, 1975;Laskey and Gurdon, 1970).…”

Section: Introductionmentioning

confidence: 99%

The influence of donor nucleus source on the outcome of zebrafish somatic cell nuclear transfer

Siripattarapravat¹,

Pinmee²,

Chang³

et al. 2010

Int. J. Dev. Biol.

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The success of nuclear reprogramming following somatic cell nuclear transfer (SCNT) is thought to depend on factors present in the egg. Little is known about the role -if any -played by the somatic cell type on the outcome of the procedure. We tested whether cells of different lineages might have different capacities for reprogramming following SCNT, comparing cells isolated from five different tissues of transgenic zebrafish for their developmental potential when used as SCNT donor cells. We used transgenic zebrafish lines expressing green fluorescence protein under an endogenous tissue-specific promoter: HGn62A-skin, HGn28A-skin, HGn8E-heart, HG21C-fin and notochord and HGn30A-hatch gland. We analyzed the efficiency of cloning, as measured by reconstructed embryos that developed up to the hatched-fry stage. Specifically, donor cells of fin and notochord origin yielded the best rate of cloned fish production. All of the other cell types used were capable of producing cloned fish, albeit with significantly lower efficiency. These results indicate that the type of zebrafish cells used for SCNT can influence the outcome of the procedure. Future epigenetic analysis of these cells will help determine specific chromatin profiles in somatic cells that have an impact on nuclear reprogramming procedures.

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“…Cloning studies in different animal species have shown that the donor cell type is one of the most important factors determining the success of cloning [1,2]. Laboratory mouse strains provide the best models for this kind of study, because they allow rigorously controlled experiments using genetically defined animals.…”

Section: Introductionmentioning

confidence: 99%

Differential Developmental Ability of Embryos Cloned from Tissue-Specific Stem Cells

et al. 2007

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Although cloning animals by somatic cell nuclear transfer is generally inefficient, the use of certain nuclear donor cell types may significantly improve or deteriorate outcomes. We evaluated whether two multipotent stem cell lines produced in vitro-neural stem cells (NSCs) and mesenchymal stem cells (MSCs)-could serve as nuclear donors for nuclear transfer cloning. Most (76%) NSCderived embryos survived the two-cell-to-four-cell transition, the stage when the major zygotic gene activation occurs. Consistent with this observation, the expression patterns of zygotically active genes were better in NSCderived embryos than in fibroblast clone embryos, which arrested at the two-cell stage more frequently. Embryo transfer experiments demonstrated that at least some of these NSC embryos had the ability to develop to term fetuses (1.6%, 3/189). In contrast, embryos reconstructed using MSCs showed a low rate of in vitro development and never underwent implantation in vivo. Chromosomal analysis of the donor MSCs revealed very frequent aneuploidy, which probably impaired the potential for development of their derived clones. This is the first demonstration that tissue-specific multipotent stem cells produced in vitro can serve as donors of nuclei for cloning mice; however, these cells may be prone to chromosomal aberrations, leading to high embryonic death rates. We found previously that hematopoietic stem cells (HSCs) are very inefficient donor cells because of their failure to activate the genes essential for embryonic development. Taken together, our data led us to conclude that tissuespecific stem cells in mice, namely NSCs, MSCs, and HSCs, exhibited marked variations in the ability to produce cloned offspring and that this ability varies according to both the epigenetic and genetic status of the original genomes.

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Donor cell differentiation, reprogramming, and cloning efficiency: Elusive or illusive correlation?

Cited by 71 publications

References 74 publications

Derivation of induced pluripotent stem cells from pig somatic cells

Derivation of induced pluripotent stem cells from pig somatic cells

The influence of donor nucleus source on the outcome of zebrafish somatic cell nuclear transfer

Differential Developmental Ability of Embryos Cloned from Tissue-Specific Stem Cells

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