OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres

David M Kristensen; John E Nielsen; Mark Kalisz; Marlene D Dalgaard; Karine Audouze; Malene E Larsen; Grete K Jacobsen; Thomas Horn; Søren Brunak; Niels E Skakkebaek; Henrik Leffers; David Møbjerg Kristensen; John E Nielsen; Mark Kalisz; Marlene D Dalgaard; Karine Marie Laure Audouze; Malene E Larsen; Grete K Jacobsen; Thomas Horn; Søren Brunak; Niels Erik Skakkebæk; Henrik Leffers

doi:10.1093/molehr/gaq008

OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres

David M Kristensen, John E Nielsen, Mark Kalisz, Marlene D Dalgaard, Karine Audouze, Malene E Larsen, Grete K Jacobsen, Thomas Horn, Søren Brunak, Niels E Skakkebaek, Henrik Leffers, David Møbjerg Kristensen, John E Nielsen, Mark Kalisz, Marlene D Dalgaard, Karine Marie Laure Audouze, Malene E Larsen, Grete K Jacobsen, Thomas Horn, Søren BrunakNiels Erik Skakkebæk, Henrik Leffers

22 Citations (Scopus)

Abstract

The transcription factor OCT4 plays a crucial role in the earliest differentiation of the mammalian embryo and in self-renewal of embryonic stem cells. However, it remains controversial whether this gene is also expressed in somatic tissues. Here, we use a combination of RT-PCR on whole and microdissected tissues, in situ hybridization, immunohistochemistry and western blotting to show that OCT4 and SOX2 together with downstream targets, UTF1 and REX1/ZFP42, are expressed in the human male urogenital tract. We further support these results by the analysis of DNA methylation of a region in the OCT4 promoter. In culture, human primary epididymal cells formed spheres that continued to express the investigated genes for at least 20 days. Transcriptomic analysis of cultured cells showed up-regulation of CD29, CD44 and CD133 that are normally associated with sphere-forming cancer stem cells. Furthermore, stimulation with retinoic acid resulted in down-regulation of OCT4 expression, however, without multilineage differentiation. Our results show that OCT4 and associated genes are expressed in somatic epithelial cells from the urogenital tract and that these cells can form spheres, a general marker of stem cell behaviour.

Original language	English
Journal	Molecular Human Reproduction
Volume	16
Issue number	11
Pages (from-to)	835-45
Number of pages	11
ISSN	1360-9947
DOIs	https://doi.org/10.1093/molehr/gaq008
Publication status	Published - 1 Nov 2010

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Kristensen, D. M., Nielsen, J. E., Kalisz, M., Dalgaard, M. D., Audouze, K., Larsen, M. E., Jacobsen, G. K., Horn, T., Brunak, S., Skakkebaek, N. E., Leffers, H., Kristensen, D. M., Nielsen, J. E., Kalisz, M., Dalgaard, M. D., Audouze, K. M. L., Larsen, M. E., Jacobsen, G. K., Horn, T., ... Leffers, H. (2010). OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres. Molecular Human Reproduction, 16(11), 835-45. https://doi.org/10.1093/molehr/gaq008

Kristensen, DM, Nielsen, JE, Kalisz, M, Dalgaard, MD, Audouze, K, Larsen, ME, Jacobsen, GK, Horn, T, Brunak, S , Skakkebaek, NE, Leffers, H, Kristensen, DM, Nielsen, JE, Kalisz, M, Dalgaard, MD, Audouze, KML, Larsen, ME, Jacobsen, GK, Horn, T, Brunak, S, Skakkebæk, NE & Leffers, H 2010, 'OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres', Molecular Human Reproduction, vol. 16, no. 11, pp. 835-45. https://doi.org/10.1093/molehr/gaq008

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title = "OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres",

abstract = "The transcription factor OCT4 plays a crucial role in the earliest differentiation of the mammalian embryo and in self-renewal of embryonic stem cells. However, it remains controversial whether this gene is also expressed in somatic tissues. Here, we use a combination of RT-PCR on whole and microdissected tissues, in situ hybridization, immunohistochemistry and western blotting to show that OCT4 and SOX2 together with downstream targets, UTF1 and REX1/ZFP42, are expressed in the human male urogenital tract. We further support these results by the analysis of DNA methylation of a region in the OCT4 promoter. In culture, human primary epididymal cells formed spheres that continued to express the investigated genes for at least 20 days. Transcriptomic analysis of cultured cells showed up-regulation of CD29, CD44 and CD133 that are normally associated with sphere-forming cancer stem cells. Furthermore, stimulation with retinoic acid resulted in down-regulation of OCT4 expression, however, without multilineage differentiation. Our results show that OCT4 and associated genes are expressed in somatic epithelial cells from the urogenital tract and that these cells can form spheres, a general marker of stem cell behaviour.",

author = "Kristensen, {David M} and Nielsen, {John E} and Mark Kalisz and Dalgaard, {Marlene D} and Karine Audouze and Larsen, {Malene E} and Jacobsen, {Grete K} and Thomas Horn and S{\o}ren Brunak and Skakkebaek, {Niels E} and Henrik Leffers and Kristensen, {David M{\o}bjerg} and Nielsen, {John E} and Mark Kalisz and Dalgaard, {Marlene D} and Audouze, {Karine Marie Laure} and Larsen, {Malene E} and Jacobsen, {Grete K} and Thomas Horn and S{\o}ren Brunak and Skakkeb{\ae}k, {Niels Erik} and Henrik Leffers",

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doi = "10.1093/molehr/gaq008",

language = "English",

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T1 - OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres

AU - Kristensen, David M

AU - Nielsen, John E

AU - Kalisz, Mark

AU - Dalgaard, Marlene D

AU - Audouze, Karine

AU - Larsen, Malene E

AU - Jacobsen, Grete K

AU - Horn, Thomas

AU - Brunak, Søren

AU - Skakkebaek, Niels E

AU - Leffers, Henrik

AU - Kristensen, David Møbjerg

AU - Nielsen, John E

AU - Kalisz, Mark

AU - Dalgaard, Marlene D

AU - Audouze, Karine Marie Laure

AU - Larsen, Malene E

AU - Jacobsen, Grete K

AU - Horn, Thomas

AU - Brunak, Søren

AU - Skakkebæk, Niels Erik

AU - Leffers, Henrik

PY - 2010/11/1

Y1 - 2010/11/1

N2 - The transcription factor OCT4 plays a crucial role in the earliest differentiation of the mammalian embryo and in self-renewal of embryonic stem cells. However, it remains controversial whether this gene is also expressed in somatic tissues. Here, we use a combination of RT-PCR on whole and microdissected tissues, in situ hybridization, immunohistochemistry and western blotting to show that OCT4 and SOX2 together with downstream targets, UTF1 and REX1/ZFP42, are expressed in the human male urogenital tract. We further support these results by the analysis of DNA methylation of a region in the OCT4 promoter. In culture, human primary epididymal cells formed spheres that continued to express the investigated genes for at least 20 days. Transcriptomic analysis of cultured cells showed up-regulation of CD29, CD44 and CD133 that are normally associated with sphere-forming cancer stem cells. Furthermore, stimulation with retinoic acid resulted in down-regulation of OCT4 expression, however, without multilineage differentiation. Our results show that OCT4 and associated genes are expressed in somatic epithelial cells from the urogenital tract and that these cells can form spheres, a general marker of stem cell behaviour.

AB - The transcription factor OCT4 plays a crucial role in the earliest differentiation of the mammalian embryo and in self-renewal of embryonic stem cells. However, it remains controversial whether this gene is also expressed in somatic tissues. Here, we use a combination of RT-PCR on whole and microdissected tissues, in situ hybridization, immunohistochemistry and western blotting to show that OCT4 and SOX2 together with downstream targets, UTF1 and REX1/ZFP42, are expressed in the human male urogenital tract. We further support these results by the analysis of DNA methylation of a region in the OCT4 promoter. In culture, human primary epididymal cells formed spheres that continued to express the investigated genes for at least 20 days. Transcriptomic analysis of cultured cells showed up-regulation of CD29, CD44 and CD133 that are normally associated with sphere-forming cancer stem cells. Furthermore, stimulation with retinoic acid resulted in down-regulation of OCT4 expression, however, without multilineage differentiation. Our results show that OCT4 and associated genes are expressed in somatic epithelial cells from the urogenital tract and that these cells can form spheres, a general marker of stem cell behaviour.

U2 - 10.1093/molehr/gaq008

DO - 10.1093/molehr/gaq008

M3 - Journal article

C2 - 20123703

SN - 1360-9947

VL - 16

SP - 835

EP - 845

JO - Molecular Human Reproduction

JF - Molecular Human Reproduction

IS - 11

ER -

OCT4 and downstream factors are expressed in human somatic urogenital epithelia and in culture of epididymal spheres

Abstract

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