[49] Derivation of embryonic stem cell lines

Abbondanzo, Susan J.; Gadi, Inder K.; Stewart, Colin L.

doi:10.1016/0076-6879(93)25052-4

Cited by 207 publications

(117 citation statements)

References 12 publications

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“…These experiments also show that more than one cell per epiblast is able to form ES cells. Because the standard practice in establishing ES lines is to pool all undifferentiated colonies obtained from a single blastocyst (10,23), at least some of the lines in general use will be polyclonal in origin. Whether this contributes to the heterogeneity of ES lines, which, according to current views, is held to arise entirely in culture (1), is clearly open to investigation now that we have shown that multiple clonal ES cell lines can be obtained from individual dissociated epiblasts.…”

Section: Discussionmentioning

confidence: 99%

The origin and efficient derivation of embryonic stem cells in the mouse

Brook

Gardner

1997

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

477

360

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By explanting tissues isolated microsurgically from implanting strain 129 mouse blastocysts individually on STO feeder cells we have established that embryonic stem (ES) cells originate from the epiblast (primitive ectoderm). Isolated early epiblasts yielded ES cell lines at a substantially higher frequency than intact blastocysts regardless of whether they were explanted whole or as strictly single-cell suspensions. When explanted from delayedimplanting 129 blastocysts, epiblasts gave lines consistently in 100% of cases. If primary embryonic fibroblasts rather than STO cells were used as feeders, germline-competent ES cell lines were obtained readily from epiblasts of delayedimplanting blastocysts of several hitherto refractory strains, particularly when recombinant leukemia inhibitory factor was included in the medium during the initial period of culture. Because lines were obtained from the nonpermissive CBA͞Ca strain at a rate of up to 56%, this approach to the derivation of germline-competent ES cell lines may not only prove generic for the mouse but also worth pursuing in other species of mammal.

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Section: Discussionmentioning

confidence: 99%

The origin and efficient derivation of embryonic stem cells in the mouse

Brook

Gardner

1997

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

477

360

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“…COS-7 and 293T cells were transfected using FuGENE6 (Roche, Indianapolis, IN, USA) following the manufacturer's instructions. We used previously reported methods (Abbondanzo et al, 1993) to isolate and culture primary MEF derived from 13.5 day embryos from Rassf1a À/À and Rassf1a þ / þ mice (Tommasi et al, 2005).…”

Section: Methodsmentioning

confidence: 99%

RASSF1A interacts with and activates the mitotic kinase Aurora-A

et al. 2008

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“…Outgrowths were disaggregated by trypsinization and grown on feeder layers for 6 d. Subsequent passaging of ES colonies was performed as described by Abbondanzo et al (1993).…”

Section: Embryonic Outgrowths and Es Cell Derivationmentioning

confidence: 99%

Oct4 distribution and level in mouse clones: consequences for pluripotency

Boiani¹,

Eckardt²,

Schöler³

et al. 2002

Genes Dev.

482

332

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Somatic cell clones often fail at a developmental stage coincident with commencement of differentiation. The transcription factor Oct4 is expressed during cleavage stages and is essential for the differentiation of the blastocyst. Oct4 expression becomes restricted to the inner cell mass and epiblast. After gastrulation Oct4 is active only in germ cells and is silent in somatic cells. Here, Oct4 and an Oct4-GFP transgene were used as markers for which gene reprogramming could be directly related to the developmental potential of somatic cell clones. Cumulus cell clones initiated Oct4 expression at the correct stage but showed an incorrect spatial expression in the majority of blastocysts. The ability of clones to form outgrowths was reduced, and the outgrowths had low or even undetectable levels of Oct4 RNA or GFP. The quality of GFP signals in blastocysts correlated with the ability to generate outgrowths that maintain GFP expression and the frequency of embryonic stem (ES) cell derivation. Abnormal Oct4 expression in clones is either directly or indirectly caused by reprogramming errors and is indicative of a general failure to reset the genetic program. The abnormal Oct4 expression may be associated with aberrant expression of other crucial developmental genes, leading to abnormalities at various embryonic stages. Regardless of other genes, the variations observed in Oct4 levels alone account for the majority of failures currently observed for somatic cell cloning.

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[49] Derivation of embryonic stem cell lines

Cited by 207 publications

References 12 publications

The origin and efficient derivation of embryonic stem cells in the mouse

The origin and efficient derivation of embryonic stem cells in the mouse

RASSF1A interacts with and activates the mitotic kinase Aurora-A

Oct4 distribution and level in mouse clones: consequences for pluripotency

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