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Rex1/Zfp42 is dispensable for pluripotency in mouse ES cells.

Masui S, Ohtsuka S, Yagi R, Takahashi K, Ko MS, Niwa H - BMC Dev. Biol. (2008)

Bottom Line: Our results clearly indicated that Rex1 function is dispensable for both the maintenance of pluripotency in ES cells and the development of embryos.However, Rex1-/- ES cells showed the defect to induce a subset of the marker genes of visceral endoderm, when differentiated as embryoid body, as found in EC cells.Rex1 should be regarded just as a marker of pluripotency without functional significance like the activity of alkaline phosphatase.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory for Pluripotent Cell Studies, RIKEN Center for Developmental Biology (CDB), 2-2-3 Minatojima-minamimachi, Kobe, Hyogo 650-0047, Japan. masui@ri.imcj.go.jp

ABSTRACT

Background: Rex1/Zfp42 has been extensively used as a marker for the undifferentiated state of pluripotent stem cells. However, its function in pluripotent stem cells including embryonic stem (ES) cells remained unclear although its involvement in visceral endoderm differentiation in F9 embryonal carcinoma (EC) cells was reported.

Results: We showed the function of Rex1 in mouse ES cells as well as in embryos using the conventional gene targeting strategy. Our results clearly indicated that Rex1 function is dispensable for both the maintenance of pluripotency in ES cells and the development of embryos. However, Rex1-/- ES cells showed the defect to induce a subset of the marker genes of visceral endoderm, when differentiated as embryoid body, as found in EC cells.

Conclusion: Rex1 should be regarded just as a marker of pluripotency without functional significance like the activity of alkaline phosphatase.

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Related in: MedlinePlus

Chimeric embryos derived from Rex1 KO ES cells. When HP4-EGFP ES cells, which were homozygotes for the mutant Rex1 allele and marked by the constitutively-active Egfp transgene, were injected into blastocysts, the embryos developed to chimeras at 12.5 dpc in which widespread contributions of GFP-positive cells were observed in fluorescent microscopic observation.
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Figure 5: Chimeric embryos derived from Rex1 KO ES cells. When HP4-EGFP ES cells, which were homozygotes for the mutant Rex1 allele and marked by the constitutively-active Egfp transgene, were injected into blastocysts, the embryos developed to chimeras at 12.5 dpc in which widespread contributions of GFP-positive cells were observed in fluorescent microscopic observation.

Mentions: To fully evaluate the differentiation ability of the Rex1-/- ES cells, we labeled these cells by introduction of the constitutively-active Egfp transgene (CAG-Egfp-IZ) and injected them into blastocysts followed by transplantation in uteri of pseudo-pregnant mice to generate chimeric embryos. As a result, we obtained the embryos with widespread contribution of the fluorescent cells derived from the Rex1-/- ES cells, indicating that these ES cells were pluripotent (Fig. 5).


Rex1/Zfp42 is dispensable for pluripotency in mouse ES cells.

Masui S, Ohtsuka S, Yagi R, Takahashi K, Ko MS, Niwa H - BMC Dev. Biol. (2008)

Chimeric embryos derived from Rex1 KO ES cells. When HP4-EGFP ES cells, which were homozygotes for the mutant Rex1 allele and marked by the constitutively-active Egfp transgene, were injected into blastocysts, the embryos developed to chimeras at 12.5 dpc in which widespread contributions of GFP-positive cells were observed in fluorescent microscopic observation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2386458&req=5

Figure 5: Chimeric embryos derived from Rex1 KO ES cells. When HP4-EGFP ES cells, which were homozygotes for the mutant Rex1 allele and marked by the constitutively-active Egfp transgene, were injected into blastocysts, the embryos developed to chimeras at 12.5 dpc in which widespread contributions of GFP-positive cells were observed in fluorescent microscopic observation.
Mentions: To fully evaluate the differentiation ability of the Rex1-/- ES cells, we labeled these cells by introduction of the constitutively-active Egfp transgene (CAG-Egfp-IZ) and injected them into blastocysts followed by transplantation in uteri of pseudo-pregnant mice to generate chimeric embryos. As a result, we obtained the embryos with widespread contribution of the fluorescent cells derived from the Rex1-/- ES cells, indicating that these ES cells were pluripotent (Fig. 5).

Bottom Line: Our results clearly indicated that Rex1 function is dispensable for both the maintenance of pluripotency in ES cells and the development of embryos.However, Rex1-/- ES cells showed the defect to induce a subset of the marker genes of visceral endoderm, when differentiated as embryoid body, as found in EC cells.Rex1 should be regarded just as a marker of pluripotency without functional significance like the activity of alkaline phosphatase.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory for Pluripotent Cell Studies, RIKEN Center for Developmental Biology (CDB), 2-2-3 Minatojima-minamimachi, Kobe, Hyogo 650-0047, Japan. masui@ri.imcj.go.jp

ABSTRACT

Background: Rex1/Zfp42 has been extensively used as a marker for the undifferentiated state of pluripotent stem cells. However, its function in pluripotent stem cells including embryonic stem (ES) cells remained unclear although its involvement in visceral endoderm differentiation in F9 embryonal carcinoma (EC) cells was reported.

Results: We showed the function of Rex1 in mouse ES cells as well as in embryos using the conventional gene targeting strategy. Our results clearly indicated that Rex1 function is dispensable for both the maintenance of pluripotency in ES cells and the development of embryos. However, Rex1-/- ES cells showed the defect to induce a subset of the marker genes of visceral endoderm, when differentiated as embryoid body, as found in EC cells.

Conclusion: Rex1 should be regarded just as a marker of pluripotency without functional significance like the activity of alkaline phosphatase.

Show MeSH
Related in: MedlinePlus