Neural differentiation of human embryonic stem cells induced by the transgene-mediated overexpression of single transcription factors

Misako Matsushita, Yuki Nakatake, Itaru Arai, Keiji Ibata, Kazuhisa Kohda, Sravan K. Goparaju, Miyako Murakami, Miki Sakota, Nana Chikazawa-Nohtomi, Shigeru Ko, Takanori Kanai, Michisuke Yuzaki, Minoru Ko

研究成果: Article

7 引用 (Scopus)

抄録

Pluripotent human embryonic stem cells (hESCs) can differentiate into multiple cell lineages, thus, providing one of the best platforms to study molecular mechanisms during cell differentiation. Recently, we have reported rapid and efficient differentiation of hESCs into functional neurons by introducing a cocktail of synthetic mRNAs encoding five transcription factors (TFs): NEUROG1, NEUROG2, NEUROG3, NEUROD1, and NEUROD2. Here we further tested a possibility that even single transcription factors, when expressed ectopically, can differentiate hESCs into neurons. To this end, we established hESC lines in which each of these TFs can be overexpressed by the doxycycline-inducible piggyBac vector. The overexpression of any of these five TFs indeed caused a rapid and rather uniform differentiation of hESCs, which were identified as neurons based on their morphologies, qRT-PCR, and immunohistochemistry. Furthermore, calcium-imaging analyses and patch clamp recordings demonstrated that these differentiated cells are electrophysiologically functional. Interestingly, neural differentiations occurred despite the cell culture conditions that rather promote the maintenance of the undifferentiated state. These results indicate that over-expression of each of these five TFs can override the pluripotency-specific gene network and force hESCs to differentiate into neurons.

元の言語English
ジャーナルBiochemical and Biophysical Research Communications
DOI
出版物ステータスAccepted/In press - 2017 5 31

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Stem cells
Transgenes
Transcription Factors
Neurons
Doxycycline
Gene Regulatory Networks
Clamping devices
Cell Lineage
Cell culture
Human Embryonic Stem Cells
Cell Differentiation
Cell Culture Techniques
Genes
Immunohistochemistry
Maintenance
Calcium
Imaging techniques
Cell Line
Polymerase Chain Reaction
Messenger RNA

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

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title = "Neural differentiation of human embryonic stem cells induced by the transgene-mediated overexpression of single transcription factors",
abstract = "Pluripotent human embryonic stem cells (hESCs) can differentiate into multiple cell lineages, thus, providing one of the best platforms to study molecular mechanisms during cell differentiation. Recently, we have reported rapid and efficient differentiation of hESCs into functional neurons by introducing a cocktail of synthetic mRNAs encoding five transcription factors (TFs): NEUROG1, NEUROG2, NEUROG3, NEUROD1, and NEUROD2. Here we further tested a possibility that even single transcription factors, when expressed ectopically, can differentiate hESCs into neurons. To this end, we established hESC lines in which each of these TFs can be overexpressed by the doxycycline-inducible piggyBac vector. The overexpression of any of these five TFs indeed caused a rapid and rather uniform differentiation of hESCs, which were identified as neurons based on their morphologies, qRT-PCR, and immunohistochemistry. Furthermore, calcium-imaging analyses and patch clamp recordings demonstrated that these differentiated cells are electrophysiologically functional. Interestingly, neural differentiations occurred despite the cell culture conditions that rather promote the maintenance of the undifferentiated state. These results indicate that over-expression of each of these five TFs can override the pluripotency-specific gene network and force hESCs to differentiate into neurons.",
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author = "Misako Matsushita and Yuki Nakatake and Itaru Arai and Keiji Ibata and Kazuhisa Kohda and Goparaju, {Sravan K.} and Miyako Murakami and Miki Sakota and Nana Chikazawa-Nohtomi and Shigeru Ko and Takanori Kanai and Michisuke Yuzaki and Minoru Ko",
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AU - Matsushita, Misako

AU - Nakatake, Yuki

AU - Arai, Itaru

AU - Ibata, Keiji

AU - Kohda, Kazuhisa

AU - Goparaju, Sravan K.

AU - Murakami, Miyako

AU - Sakota, Miki

AU - Chikazawa-Nohtomi, Nana

AU - Ko, Shigeru

AU - Kanai, Takanori

AU - Yuzaki, Michisuke

AU - Ko, Minoru

PY - 2017/5/31

Y1 - 2017/5/31

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AB - Pluripotent human embryonic stem cells (hESCs) can differentiate into multiple cell lineages, thus, providing one of the best platforms to study molecular mechanisms during cell differentiation. Recently, we have reported rapid and efficient differentiation of hESCs into functional neurons by introducing a cocktail of synthetic mRNAs encoding five transcription factors (TFs): NEUROG1, NEUROG2, NEUROG3, NEUROD1, and NEUROD2. Here we further tested a possibility that even single transcription factors, when expressed ectopically, can differentiate hESCs into neurons. To this end, we established hESC lines in which each of these TFs can be overexpressed by the doxycycline-inducible piggyBac vector. The overexpression of any of these five TFs indeed caused a rapid and rather uniform differentiation of hESCs, which were identified as neurons based on their morphologies, qRT-PCR, and immunohistochemistry. Furthermore, calcium-imaging analyses and patch clamp recordings demonstrated that these differentiated cells are electrophysiologically functional. Interestingly, neural differentiations occurred despite the cell culture conditions that rather promote the maintenance of the undifferentiated state. These results indicate that over-expression of each of these five TFs can override the pluripotency-specific gene network and force hESCs to differentiate into neurons.

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