TY - JOUR
T1 - Reprogramming of chimpanzee fibroblasts into a multipotent cancerous but not fully pluripotent state by transducing iPSC factors in 2i/LIF culture
AU - Lin, Zachary Yu Ching
AU - Nakai, Risako
AU - Hirai, Hirohisa
AU - Kozuka, Daiki
AU - Katayama, Seiya
AU - Nakamura, Shin ichiro
AU - Okada, Sawako
AU - Kitajima, Ryunosuke
AU - Imai, Hiroo
AU - Okano, Hideyuki
AU - Imamura, Masanori
N1 - Funding Information:
We thank the staff at Sapporo Maruyama Zoo, Tobu Zoo, Tama Zoological Park, Beppu Cable Rakutenchi, Primate Research Institute, and the Great Ape Information Network (GAIN) for chimpanzee materials, and Koji Nishihara, Yohei Bamba, Jia-Ni Lin, Yohei Okada, Hirotaka James Okano, Haruka Ito, Yuriko Hirai, and Kaori Yasutake for technical assistance and other support. We also thank Mitchell Arico from Edanz Group (www.edanzediting.com/ac) for editing a draft of this manuscript. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT); the Ministry of Health, Labour, and Welfare; the Japan Society for the Promotion of Science (JSPS); the National Institute of Biomedical Innovation and the Strategic Research Program for Brain Sciences from MEXT; the Funding Program for World-leading Innovative R&D in Science and Technology (FIRST); a Keio University Grant-in-Aid for the Encouragement of Young Medical Scientists; Leave a Nest Grants (Life Technologies Japan Award, On-chip Biotechnologies Award, SCREEN Holdings Award, L-RAD Award); Interuniversity Bio-Backup Project for Basic Biology; the Brain Sciences Project of the Center for Novel Science Initiatives (CNTI), National Institutes of Natural Sciences (NINS); Aichi Cancer Research Foundation; Takeda Science Foundation; The Kyoto University Foundation; The Hori Science and Arts Foundation; the Cooperation Research Program of the Primate Research Institute, Kyoto University; Extramural Collaborative Research Grant of Cancer Research Institute, Kanazawa University; and the Cooperative Study Program of National Institute for Physiological Sciences. Z.Y.L was supported by the Otsuka Toshimi foundation. R.N. was supported by the DAIKO Foundation and the Kyoto University Graduate School of Science Fund.
Funding Information:
We thank the staff at Sapporo Maruyama Zoo, Tobu Zoo, Tama Zoological Park, Beppu Cable Rakutenchi, Primate Research Institute, and the Great Ape Information Network (GAIN) for chimpanzee materials, and Koji Nishihara, Yohei Bamba, Jia-Ni Lin, Yohei Okada, Hirotaka James Okano, Haruka Ito, Yuriko Hirai, and Kaori Yasutake for technical assistance and other support. We also thank Mitchell Arico from Edanz Group ( www.edanzediting.com/ac ) for editing a draft of this manuscript. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT) ; the Ministry of Health, Labour, and Welfare ; the Japan Society for the Promotion of Science (JSPS) ; the National Institute of Biomedical Innovation and the Strategic Research Program for Brain Sciences from MEXT ; the Funding Program for World-leading Innovative R&D in Science and Technology (FIRST) ; a Keio University Grant-in-Aid for the Encouragement of Young Medical Scientists; Leave a Nest Grants (Life Technologies Japan Award, On-chip Biotechnologies Award, SCREEN Holdings Award, L-RAD Award); Interuniversity Bio-Backup Project for Basic Biology; the Brain Sciences Project of the Center for Novel Science Initiatives (CNTI), National Institutes of Natural Sciences (NINS); Aichi Cancer Research Foundation; Takeda Science Foundation; The Kyoto University Foundation; The Hori Science and Arts Foundation; the Cooperation Research Program of the Primate Research Institute, Kyoto University; Extramural Collaborative Research Grant of Cancer Research Institute, Kanazawa University; and the Cooperative Study Program of National Institute for Physiological Sciences. Z.Y.L was supported by the Otsuka Toshimi foundation . R.N. was supported by the DAIKO Foundation and the Kyoto University Graduate School of Science Fund .
Publisher Copyright:
© 2020 International Society of Differentiation
PY - 2020/3/1
Y1 - 2020/3/1
N2 - To induce and maintain naïve pluripotency in mouse embryonic and induced pluripotent stem cells (ESCs/iPSCs), chemically defined N2B27 medium with PD0325901, CHIR99021, and leukemia inhibitory factor (2i/LIF) is a classic and simple condition. However, this method cannot be simply extrapolated to human ESCs/iPSCs that are principally stabilized in primed pluripotency and become primitive neuroepithelium-like cells in N2B27+2i/LIF culture. Here, we assessed iPSC reprogramming of fibroblasts from chimpanzee, our closest living relative, in N2B27+2i/LIF culture. Under this condition, chimpanzee cells formed alkaline phosphatase-positive dome-shaped colonies. The colony-forming cells could be stably expanded by serial passaging without a ROCK inhibitor. However, their gene expression was distinct from iPSCs and neuroepithelium. They expressed the OCT3/4 transgene and a subset of transcripts associated with pluripotency, mesenchymal-epithelial transition, and neural crest formation. These cells exhibited a differentiation potential into the three germ layers in vivo and in vitro. The current study demonstrated that iPSC reprogramming in N2B27+2i/LIF culture converted chimpanzee fibroblasts into a multipotent cancerous state with unique gene expression, but not fully pluripotent stem cells.
AB - To induce and maintain naïve pluripotency in mouse embryonic and induced pluripotent stem cells (ESCs/iPSCs), chemically defined N2B27 medium with PD0325901, CHIR99021, and leukemia inhibitory factor (2i/LIF) is a classic and simple condition. However, this method cannot be simply extrapolated to human ESCs/iPSCs that are principally stabilized in primed pluripotency and become primitive neuroepithelium-like cells in N2B27+2i/LIF culture. Here, we assessed iPSC reprogramming of fibroblasts from chimpanzee, our closest living relative, in N2B27+2i/LIF culture. Under this condition, chimpanzee cells formed alkaline phosphatase-positive dome-shaped colonies. The colony-forming cells could be stably expanded by serial passaging without a ROCK inhibitor. However, their gene expression was distinct from iPSCs and neuroepithelium. They expressed the OCT3/4 transgene and a subset of transcripts associated with pluripotency, mesenchymal-epithelial transition, and neural crest formation. These cells exhibited a differentiation potential into the three germ layers in vivo and in vitro. The current study demonstrated that iPSC reprogramming in N2B27+2i/LIF culture converted chimpanzee fibroblasts into a multipotent cancerous state with unique gene expression, but not fully pluripotent stem cells.
KW - Chimpanzee
KW - Pluripotency
KW - Reprogramming
KW - iPSC
UR - http://www.scopus.com/inward/record.url?scp=85078993858&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85078993858&partnerID=8YFLogxK
U2 - 10.1016/j.diff.2020.01.002
DO - 10.1016/j.diff.2020.01.002
M3 - Article
C2 - 32045848
AN - SCOPUS:85078993858
SN - 0301-4681
VL - 112
SP - 67
EP - 76
JO - Differentiation
JF - Differentiation
ER -
