Improved Generation of Germline‐Competent Embryonic Stem Cell Lines from Inbred Mouse Strains

Schoonjans, Luc; Kreemers, Veerle; Danloy, Sophie; Moreadith, Randall W.; Laroche, Y; Collen, Désiré

doi:10.1634/stemcells.21-1-90

Cited by 100 publications

(75 citation statements)

References 11 publications

(13 reference statements)

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“…and A.N., unpublished data) or with very high levels of ES cell contribution when the cells were injected under the zona pellucida of morula stage diploid embryos (personal communication). Many factors such as the genetic background of the host embryo, the method of chimera generation and the culture conditions of ES cells can influence the level of contribution of ES cells in chimeras and the behavior of their derivatives in adult tissues and organs (27). These findings suggest that heterozygosity (F 1 hybrid origin) of ES cells maybe sufficient but not necessary for this phenomenon.…”

Section: Discussionmentioning

confidence: 99%

Developmental and adult phenotyping directly from mutant embryonic stem cells

George

Gertsenstein

Vintersten

et al. 2007

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

219

196

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Tetraploid embryo complementation assay has shown that mouse ES cells alone are capable of supporting embryonic development and adult life of mice. Newly established F 1 hybrid ES cells allow the production of ES cell-derived animals at a high enough efficiency to directly make ES cell-based genetics feasible. Here we report the establishment and characterization of 12 new F1 hybrid ES cell lines and the use of one of the best (G4) in a gain-and loss-of-function genetic study, where the in vivo phenotypes were assessed directly from ES cell-derived embryos. We found the generation of G4 ES cell-derived animals to be very efficient. Furthermore, even after two consecutive rounds of genetic modifications, the majority of transgenic lines retained the original potential of the parental lines; with 10 -40% of chimeras producing ES cell-derived animals/embryos. Using these genetically altered ES cells, this success rate, in most cases, permitted the derivation of a sufficient number of mutants for initial phenotypic analyses only a few weeks after the establishment of the cell lines. Although the experimental design has to take into account a moderate level of uncontrolled damage on ES cell lines, our proofof-principle experiment provides useful data to assist future designs harnessing the power of this technology to accelerate our understanding of gene function.hybrid ͉ tetraploid complementation assay ͉ vasculogenesis ͉ ES cells T he advent of mouse ES cells (1, 2) has revolutionized the genetic approaches addressing gene function. It helped transform the mouse into the ultimate mammalian model system with significant relevance to human biology. Mutating all of the genes in the mouse has become not only feasible but also a substantial international project supported by the European Union, the National Institutes of Health, and Genome Canada (3). Before this effort, thousands of ES cell lines had already been created, each representing a specific mutation, and many more are currently under way. Introduction of these cells back into the mouse through germ-line transmission is a time-, labor-, and cost-intensive endeavor, followed by the tedious task of phenotypic analyses. Therefore, an assay system, which could accelerate the production of mutant embryos or animals and at the same time provide information about in vivo phenotypes for elucidating gene function in normal developmental and pathological processes, would be of major value.Two specific properties of mouse ES cells render them exceptional tools for genetic research: (i) virtually unlimited proliferation capacity and (ii) pluripotent developmental potential. Their proliferation capability leads to the production of large number of cells and therefore the occurrence and the identification of very rare events, such as homologous recombination or gene trap insertion. The pluripotent developmental potential of ES cells allows them to contribute to the germ line when reintroduced into an embryonic environment through chimera formation. However, an additional pro...

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Section: Discussionmentioning

confidence: 99%

Developmental and adult phenotyping directly from mutant embryonic stem cells

George

Gertsenstein

Vintersten

et al. 2007

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

219

196

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“…Pdgfc +/À Pdgfa +/À mice were intercrossed, and the 2.5-d.p.c. blastocysts were flushed and then cultured in ES culture medium as described 30 . Inner cell mass outgrowths were then disaggregated into small, multicellular clumps that were cultured further until ES cell colonies developed.…”

Section: Methodsmentioning

confidence: 99%

A specific requirement for PDGF-C in palate formation and PDGFR-α signaling

et al. 2004

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“…Recently, a few FVB-ES cell lines have been reported. However, only one wildtype ES cell line was described germ line-competent, 33 whereas STAT3 overexpression in the inner cell mass achieved the germ line competency of FVB strain-derived ES cells. 34 These data may suggest that inbred FVB strain is optimal to test the reliability or efficacy of new reprogramming method.…”

Section: Protein-based Reprogramming Is Neither By the Contamination mentioning

confidence: 99%

Induction of pluripotent stem cells from adult somatic cells by protein-based reprogramming without genetic manipulation

Cho

Lee²,

Kwon

et al. 2010

Blood

218

203

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The concept of reprogramming of somatic cells has opened a new era in regenerative medicine. Transduction of defined factors has successfully achieved pluripotency. However, during the generation process of induced pluripotent stem (iPS) cells, genetic manipulation of certain factors may cause tumorigenicity, which limits further application. We report that that a single transfer of embryonic stem (ES) cell-derived proteins into primarily cultured adult mouse fibroblasts, rather than repeated transfer or prolonged exposure to materials, can achieve full reprogramming up to the pluripotent state without the forced expression of ectopic transgenes. During the process, gene expression and epigenetic status were converted from somatic to ES-equivalent status. We verified that protein-based reprogramming was neither by the contamination of protein do-

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Improved Generation of Germline‐Competent Embryonic Stem Cell Lines from Inbred Mouse Strains

Cited by 100 publications

References 11 publications

Developmental and adult phenotyping directly from mutant embryonic stem cells

Developmental and adult phenotyping directly from mutant embryonic stem cells

A specific requirement for PDGF-C in palate formation and PDGFR-α signaling

Induction of pluripotent stem cells from adult somatic cells by protein-based reprogramming without genetic manipulation

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