Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species

Sho Yoshimatsu; Mayutaka Nakajima; Aozora Iguchi; Tsukasa Sanosaka; Tsukika Sato; Mari Nakamura; Ryusuke Nakajima; Eri Arai; Mitsuru Ishikawa; Kent Imaizumi; Hirotaka Watanabe; Junko Okahara; Toshiaki Noce; Yuta Takeda; Erika Sasaki; Rüdiger Behr; Kazuya Edamura; Seiji Shiozawa; Hideyuki Okano

doi:10.1016/j.stemcr.2021.03.002

Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species

Sho Yoshimatsu, Mayutaka Nakajima, Aozora Iguchi, Tsukasa Sanosaka, Tsukika Sato, Mari Nakamura, Ryusuke Nakajima, Eri Arai, Mitsuru Ishikawa, Kent Imaizumi, Hirotaka Watanabe, Junko Okahara, Toshiaki Noce, Yuta Takeda, Erika Sasaki, Rüdiger Behr, Kazuya Edamura, Seiji Shiozawa, Hideyuki Okano

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31 Citations (Scopus)

Abstract

Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers and germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem cell-based therapy, gene-modified animal production, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrate the iPSC derivation from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the fibroblasts temporarily acquired a neural stem cell-like state during the reprogramming. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem cell-based research using various animals in Mammalia.

Original language	English
Pages (from-to)	754-770
Number of pages	17
Journal	Stem cell reports
Volume	16
Issue number	4
DOIs	https://doi.org/10.1016/j.stemcr.2021.03.002
Publication status	Published - 2021 Apr 13

Keywords

dog
episomal vector
induced pluripotent stem cell
mammalian
marmoset
neural stem cell
pig
primordial germ cell
reprogramming

ASJC Scopus subject areas

Biochemistry
Genetics
Developmental Biology
Cell Biology

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10.1016/j.stemcr.2021.03.002

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Yoshimatsu, S., Nakajima, M., Iguchi, A., Sanosaka, T., Sato, T., Nakamura, M., Nakajima, R., Arai, E., Ishikawa, M., Imaizumi, K., Watanabe, H., Okahara, J., Noce, T., Takeda, Y., Sasaki, E., Behr, R., Edamura, K., Shiozawa, S., & Okano, H. (2021). Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species. Stem cell reports, 16(4), 754-770. https://doi.org/10.1016/j.stemcr.2021.03.002

Yoshimatsu, S, Nakajima, M, Iguchi, A, Sanosaka, T, Sato, T, Nakamura, M, Nakajima, R, Arai, E, Ishikawa, M, Imaizumi, K, Watanabe, H, Okahara, J, Noce, T, Takeda, Y, Sasaki, E, Behr, R, Edamura, K, Shiozawa, S & Okano, H 2021, 'Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species', Stem cell reports, vol. 16, no. 4, pp. 754-770. https://doi.org/10.1016/j.stemcr.2021.03.002

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title = "Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species",

abstract = "Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers and germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem cell-based therapy, gene-modified animal production, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrate the iPSC derivation from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the fibroblasts temporarily acquired a neural stem cell-like state during the reprogramming. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem cell-based research using various animals in Mammalia.",

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author = "Sho Yoshimatsu and Mayutaka Nakajima and Aozora Iguchi and Tsukasa Sanosaka and Tsukika Sato and Mari Nakamura and Ryusuke Nakajima and Eri Arai and Mitsuru Ishikawa and Kent Imaizumi and Hirotaka Watanabe and Junko Okahara and Toshiaki Noce and Yuta Takeda and Erika Sasaki and R{\"u}diger Behr and Kazuya Edamura and Seiji Shiozawa and Hideyuki Okano",

note = "Funding Information: We especially thank Drs. Takashi Sasaki, Jun Kohyama, Ayako Y. Murayama, and Ms. Kanae Ohtsu (at Keio University), Drs. Hirotaka Kondo and Hisashi Shibuya (at Nihon University). We also thank all the laboratory members of H.O. and K.E. for their encouragement and generous support for the current study. This study was performed under the Strategic Research Program for Brain Sciences and Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) of the the AMED (JP20dm0207001 to H.O.), KAKENHI 19J12871 and 20K22660 (to S.Y.), 20H03156 (to K.E.), and internal budgets from Keio University, including the Program for the Advancement of Research in Core Projects on Longevity of the Keio University Global Research Institute from Keio University (to H.O.). Publisher Copyright: {\textcopyright} 2021 The Authors",

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T1 - Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species

AU - Yoshimatsu, Sho

AU - Nakajima, Mayutaka

AU - Iguchi, Aozora

AU - Sanosaka, Tsukasa

AU - Sato, Tsukika

AU - Nakamura, Mari

AU - Nakajima, Ryusuke

AU - Arai, Eri

AU - Ishikawa, Mitsuru

AU - Imaizumi, Kent

AU - Watanabe, Hirotaka

AU - Okahara, Junko

AU - Noce, Toshiaki

AU - Takeda, Yuta

AU - Sasaki, Erika

AU - Behr, Rüdiger

AU - Edamura, Kazuya

AU - Shiozawa, Seiji

AU - Okano, Hideyuki

N1 - Funding Information: We especially thank Drs. Takashi Sasaki, Jun Kohyama, Ayako Y. Murayama, and Ms. Kanae Ohtsu (at Keio University), Drs. Hirotaka Kondo and Hisashi Shibuya (at Nihon University). We also thank all the laboratory members of H.O. and K.E. for their encouragement and generous support for the current study. This study was performed under the Strategic Research Program for Brain Sciences and Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) of the the AMED (JP20dm0207001 to H.O.), KAKENHI 19J12871 and 20K22660 (to S.Y.), 20H03156 (to K.E.), and internal budgets from Keio University, including the Program for the Advancement of Research in Core Projects on Longevity of the Keio University Global Research Institute from Keio University (to H.O.). Publisher Copyright: © 2021 The Authors

PY - 2021/4/13

Y1 - 2021/4/13

N2 - Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers and germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem cell-based therapy, gene-modified animal production, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrate the iPSC derivation from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the fibroblasts temporarily acquired a neural stem cell-like state during the reprogramming. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem cell-based research using various animals in Mammalia.

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Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species

Abstract

Keywords

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