Direct reprogramming of rat neural precursor cells and fibroblasts into pluripotent stem cells

Mi Yoon Chang, Dohoon Kim, Chun Hyung Kim, hoonchul kang, Eungi Yang, Jung Il Moon, Sanghyeok Ko, Junpil Park, Kyung Soon Park, Kyung Ah Lee, Dong Youn Hwang, Young Chung, Robert Lanza, Kwang Soo Kim

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease. Copyright:

Original languageEnglish
Article numbere9838
JournalPLoS One
Volume5
Issue number3
DOIs
Publication statusPublished - 2010 Dec 1

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Pluripotent Stem Cells
Fibroblasts
Induced Pluripotent Stem Cells
Stem cells
fibroblasts
stem cells
Rats
rats
cells
human physiology
myc Genes
embryonic stem cells
Physiology
Embryonic Stem Cells
human diseases
Genes
researchers
Research Personnel
Glycogen Synthase Kinase 3
induced pluripotent stem cells

All Science Journal Classification (ASJC) codes

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

Cite this

Chang, Mi Yoon ; Kim, Dohoon ; Kim, Chun Hyung ; kang, hoonchul ; Yang, Eungi ; Moon, Jung Il ; Ko, Sanghyeok ; Park, Junpil ; Park, Kyung Soon ; Lee, Kyung Ah ; Hwang, Dong Youn ; Chung, Young ; Lanza, Robert ; Kim, Kwang Soo. / Direct reprogramming of rat neural precursor cells and fibroblasts into pluripotent stem cells. In: PLoS One. 2010 ; Vol. 5, No. 3.
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abstract = "Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease. Copyright:",
author = "Chang, {Mi Yoon} and Dohoon Kim and Kim, {Chun Hyung} and hoonchul kang and Eungi Yang and Moon, {Jung Il} and Sanghyeok Ko and Junpil Park and Park, {Kyung Soon} and Lee, {Kyung Ah} and Hwang, {Dong Youn} and Young Chung and Robert Lanza and Kim, {Kwang Soo}",
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Chang, MY, Kim, D, Kim, CH, kang, H, Yang, E, Moon, JI, Ko, S, Park, J, Park, KS, Lee, KA, Hwang, DY, Chung, Y, Lanza, R & Kim, KS 2010, 'Direct reprogramming of rat neural precursor cells and fibroblasts into pluripotent stem cells', PLoS One, vol. 5, no. 3, e9838. https://doi.org/10.1371/journal.pone.0009838

Direct reprogramming of rat neural precursor cells and fibroblasts into pluripotent stem cells. / Chang, Mi Yoon; Kim, Dohoon; Kim, Chun Hyung; kang, hoonchul; Yang, Eungi; Moon, Jung Il; Ko, Sanghyeok; Park, Junpil; Park, Kyung Soon; Lee, Kyung Ah; Hwang, Dong Youn; Chung, Young; Lanza, Robert; Kim, Kwang Soo.

In: PLoS One, Vol. 5, No. 3, e9838, 01.12.2010.

Research output: Contribution to journalArticle

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T1 - Direct reprogramming of rat neural precursor cells and fibroblasts into pluripotent stem cells

AU - Chang, Mi Yoon

AU - Kim, Dohoon

AU - Kim, Chun Hyung

AU - kang, hoonchul

AU - Yang, Eungi

AU - Moon, Jung Il

AU - Ko, Sanghyeok

AU - Park, Junpil

AU - Park, Kyung Soon

AU - Lee, Kyung Ah

AU - Hwang, Dong Youn

AU - Chung, Young

AU - Lanza, Robert

AU - Kim, Kwang Soo

PY - 2010/12/1

Y1 - 2010/12/1

N2 - Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease. Copyright:

AB - Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease. Copyright:

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