Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors

Takamasa Kawaguchi, Dooseon Cho, Masafumi Hayashi, Tomoyuki Tsukiyama, Koji Kimura, Shuichi Matsuyama, Naojiro Minami, Masayasu Yamada, Hiroshi Imai

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3/4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast- related (CDX2, ELF5, ERRβ, and IFN-t) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy- related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.

Original languageEnglish
Article numbere0167550
JournalPLoS One
Volume11
Issue number12
DOIs
Publication statusPublished - Dec 1 2016

Fingerprint

trophoblast
doxycycline
Doxycycline
Trophoblasts
Cells
cell lines
Cell Line
cattle
Stem cells
Genes
cells
Amnion
amnion
Cell Lineage
Balloons
Transcription Factors
Induced Pluripotent Stem Cells
Specifications
Stem Cells
stem cells

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Kawaguchi, T., Cho, D., Hayashi, M., Tsukiyama, T., Kimura, K., Matsuyama, S., ... Imai, H. (2016). Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors. PLoS One, 11(12), [e0167550]. https://doi.org/10.1371/journal.pone.0167550

Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors. / Kawaguchi, Takamasa; Cho, Dooseon; Hayashi, Masafumi; Tsukiyama, Tomoyuki; Kimura, Koji; Matsuyama, Shuichi; Minami, Naojiro; Yamada, Masayasu; Imai, Hiroshi.

In: PLoS One, Vol. 11, No. 12, e0167550, 01.12.2016.

Research output: Contribution to journalArticle

Kawaguchi, T, Cho, D, Hayashi, M, Tsukiyama, T, Kimura, K, Matsuyama, S, Minami, N, Yamada, M & Imai, H 2016, 'Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors', PLoS One, vol. 11, no. 12, e0167550. https://doi.org/10.1371/journal.pone.0167550
Kawaguchi, Takamasa ; Cho, Dooseon ; Hayashi, Masafumi ; Tsukiyama, Tomoyuki ; Kimura, Koji ; Matsuyama, Shuichi ; Minami, Naojiro ; Yamada, Masayasu ; Imai, Hiroshi. / Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors. In: PLoS One. 2016 ; Vol. 11, No. 12.
@article{781eed1ff3e645f8a582119f5fd78bd6,
title = "Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors",
abstract = "Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3/4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast- related (CDX2, ELF5, ERRβ, and IFN-t) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy- related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.",
author = "Takamasa Kawaguchi and Dooseon Cho and Masafumi Hayashi and Tomoyuki Tsukiyama and Koji Kimura and Shuichi Matsuyama and Naojiro Minami and Masayasu Yamada and Hiroshi Imai",
year = "2016",
month = "12",
day = "1",
doi = "10.1371/journal.pone.0167550",
language = "English",
volume = "11",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "12",

}

TY - JOUR

T1 - Derivation of induced trophoblast cell lines in cattle by doxycycline-inducible piggyBac vectors

AU - Kawaguchi, Takamasa

AU - Cho, Dooseon

AU - Hayashi, Masafumi

AU - Tsukiyama, Tomoyuki

AU - Kimura, Koji

AU - Matsuyama, Shuichi

AU - Minami, Naojiro

AU - Yamada, Masayasu

AU - Imai, Hiroshi

PY - 2016/12/1

Y1 - 2016/12/1

N2 - Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3/4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast- related (CDX2, ELF5, ERRβ, and IFN-t) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy- related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.

AB - Trophectoderm lineage specification is one of the earliest differentiation events in mammalian development. The trophoblast lineage, which is derived from the trophectoderm, mediates implantation and placental formation. However, the processes involved in trophoblastic differentiation and placental formation in cattle remain unclear due to interspecies differences when compared with other model systems and the small repertoire of available trophoblast cell lines. Here, we describe the generation of trophoblast cell lines (biTBCs) from bovine amnion-derived cells (bADCs) using an induced pluripotent stem cell technique. bADCs were introduced with piggyBac vectors containing doxycycline (Dox)-inducible transcription factors (Oct3/4(POU5F1), Sox2, Klf4, and c-Myc). Colonies that appeared showed a flattened epithelial-like morphology similar to cobblestones, had a more definite cell boundary between cells, and frequently formed balloon-like spheroids similar to trophoblastic vesicles (TVs). biTBCs were propagated for over 60 passages and expressed trophoblast- related (CDX2, ELF5, ERRβ, and IFN-t) and pluripotency-related genes (endogenous OCT3/4, SOX2, KLF4, and c-MYC). Furthermore, when biTBCs were induced to differentiate by removing Dox from culture, they formed binucleate cells and began to express pregnancy- related genes (PL, PRP1, and PAG1). This is the first report demonstrating that the induction of pluripotency in bovine amniotic cells allows the generation of trophoblastic cell lines that possess trophoblast stem cell-like characteristics and have the potential to differentiate into the extra-embryonic cell lineage. These cell lines can be a new cell source as a model for studying trophoblast cell lineages and implantation processes in cattle.

UR - http://www.scopus.com/inward/record.url?scp=85000797873&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85000797873&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0167550

DO - 10.1371/journal.pone.0167550

M3 - Article

C2 - 27907214

AN - SCOPUS:85000797873

VL - 11

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 12

M1 - e0167550

ER -