Inactivation of the FGF-4 gene in embryonic stem cells alters the growth and/or the survival of their early differentiated progeny

Phillip J. Wilder, David Lee Kelly, Kristen Brigman, Cynthia L. Peterson, Tamara Nowling, Qing Sheng Gao, Rodney D McComb, Mario R. Capecchi, A Angie Rizzino

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Abstract

Previous studies have shown that early mouse embryos with both FGF-4 alleles inactivated are developmentally arrested shortly after implantation. To understand the roles of FGF-4 during early development, we prepared genetically engineered embryonic stem (ES) cells, which are unable to produce FGF-4. Specifically, we describe the isolation and characterization of ES cells with both FGF-4 alleles inactivated. The FGF-4(-/-) ES cells do not require FGF-4 to proliferate m vitro, and addition of FGF-4 to the medium has little or no effect on their growth. Thus, FGF-4 does not appear to act as an autocrine growth factor for cultured ES cells. We also demonstrate that FGF- 4(-/-) ES cells, like their unmodified counterparts, are capable of forming highly complex tumors in syngeneic mice composed of a wide range of differentiated cells types, including neural tissue, glandular epithelium, and muscle. In addition, we demonstrate that the FGF-4(-/-) ES cells can differentiate m vitro after exposure to retinoic acid; however, the growth and/or survival of the differentiated cells is severely compromised. Importantly, addition of FGF-4 to the culture medium dramatically increases the number of differentiated cells derived from the FGF-4(-/-) ES cells, in particular cells with many of the properties of parietal extraembryonic endoderm. Finally, we demonstrate that there are differences in the RNA profiles expressed by the differentiated progeny formed in vitro from FGF- 4(-/-) ES cells and FGF-4(+/+) ES cells when they are cultured with FGF-4. Taken together, the studies described in this report indicate that certain lineages formed in vitro are affected by the inactivation of the FGF-4 gene, in particular specific cells that form during the initial stage of ES cell differentiation. Thus, ES cells with both FGF-4 alleles inactivated should shed light on the important roles of FGF-4 during the early stages of mammalian development and help determine why FGF-4(-/-) embryos die shortly after implantation.

Original languageEnglish (US)
Pages (from-to)614-629
Number of pages16
JournalDevelopmental Biology
Volume192
Issue number2
DOIs
StatePublished - Dec 15 1997

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Embryonic Stem Cells
Growth
Genes
Alleles
Embryonic Structures
Endoderm
Tretinoin
Culture Media
Cell Differentiation
Cell Survival
Intercellular Signaling Peptides and Proteins
Epithelium
Cell Count
RNA
Muscles

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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Inactivation of the FGF-4 gene in embryonic stem cells alters the growth and/or the survival of their early differentiated progeny. / Wilder, Phillip J.; Kelly, David Lee; Brigman, Kristen; Peterson, Cynthia L.; Nowling, Tamara; Gao, Qing Sheng; McComb, Rodney D; Capecchi, Mario R.; Rizzino, A Angie.

In: Developmental Biology, Vol. 192, No. 2, 15.12.1997, p. 614-629.

Research output: Contribution to journalArticle

Wilder, Phillip J. ; Kelly, David Lee ; Brigman, Kristen ; Peterson, Cynthia L. ; Nowling, Tamara ; Gao, Qing Sheng ; McComb, Rodney D ; Capecchi, Mario R. ; Rizzino, A Angie. / Inactivation of the FGF-4 gene in embryonic stem cells alters the growth and/or the survival of their early differentiated progeny. In: Developmental Biology. 1997 ; Vol. 192, No. 2. pp. 614-629.
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AU - Kelly, David Lee

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AU - Peterson, Cynthia L.

AU - Nowling, Tamara

AU - Gao, Qing Sheng

AU - McComb, Rodney D

AU - Capecchi, Mario R.

AU - Rizzino, A Angie

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AB - Previous studies have shown that early mouse embryos with both FGF-4 alleles inactivated are developmentally arrested shortly after implantation. To understand the roles of FGF-4 during early development, we prepared genetically engineered embryonic stem (ES) cells, which are unable to produce FGF-4. Specifically, we describe the isolation and characterization of ES cells with both FGF-4 alleles inactivated. The FGF-4(-/-) ES cells do not require FGF-4 to proliferate m vitro, and addition of FGF-4 to the medium has little or no effect on their growth. Thus, FGF-4 does not appear to act as an autocrine growth factor for cultured ES cells. We also demonstrate that FGF- 4(-/-) ES cells, like their unmodified counterparts, are capable of forming highly complex tumors in syngeneic mice composed of a wide range of differentiated cells types, including neural tissue, glandular epithelium, and muscle. In addition, we demonstrate that the FGF-4(-/-) ES cells can differentiate m vitro after exposure to retinoic acid; however, the growth and/or survival of the differentiated cells is severely compromised. Importantly, addition of FGF-4 to the culture medium dramatically increases the number of differentiated cells derived from the FGF-4(-/-) ES cells, in particular cells with many of the properties of parietal extraembryonic endoderm. Finally, we demonstrate that there are differences in the RNA profiles expressed by the differentiated progeny formed in vitro from FGF- 4(-/-) ES cells and FGF-4(+/+) ES cells when they are cultured with FGF-4. Taken together, the studies described in this report indicate that certain lineages formed in vitro are affected by the inactivation of the FGF-4 gene, in particular specific cells that form during the initial stage of ES cell differentiation. Thus, ES cells with both FGF-4 alleles inactivated should shed light on the important roles of FGF-4 during the early stages of mammalian development and help determine why FGF-4(-/-) embryos die shortly after implantation.

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