TY - JOUR
T1 - Improved Sendai viral system for reprogramming to naive pluripotency
AU - Kunitomi, Akira
AU - Hirohata, Ryoko
AU - Arreola, Vanessa
AU - Osawa, Mitsujiro
AU - Kato, Tomoaki M.
AU - Nomura, Masaki
AU - Kawaguchi, Jitsutaro
AU - Hara, Hiroto
AU - Kusano, Kohji
AU - Takashima, Yasuhiro
AU - Takahashi, Kazutoshi
AU - Fukuda, Keiichi
AU - Takasu, Naoko
AU - Yamanaka, Shinya
N1 - Funding Information:
We are grateful to M. Iwasaki and K. Tomoda for sharing data and discussions prior to publication; M. Saito, A. Niwa, and M. Nakagawa for providing valuable experimental equipment; T. Okubo for technical assistance; and P. Karagiannis and K. Claiborn for critical reading of this manuscript. This work was supported by the Core Center for iPS Cell Research , Research Center Network for Realization of Regenerative Medicine , AMED under grant number JP21bm0104001 ; iPS Cell Research Fund; and JSPS KAKENHI under grant numbers 16K19429 and 18K15846 . This work was also supported by funding from Mr. Hiroshi Mikitani, Mr. Marc Benioff, the L. K. Whittier Foundation , the Roddenberry Foundation , the Gladstone Institutes , the National Heart, Lung, and Blood Institute (NHLBI), and National Institutes of Health (NIH) ( U01-HL100406 , U01-HL098179 , R01-HL130533 , and R01-HL135358 ). Gladstone Institutes received support from National Center for Research Resources grant RR18928-01 .
Funding Information:
We are grateful to M. Iwasaki and K. Tomoda for sharing data and discussions prior to publication; M. Saito, A. Niwa, and M. Nakagawa for providing valuable experimental equipment; T. Okubo for technical assistance; and P. Karagiannis and K. Claiborn for critical reading of this manuscript. This work was supported by the Core Center for iPS Cell Research, Research Center Network for Realization of Regenerative Medicine, AMED under grant number JP21bm0104001; iPS Cell Research Fund; and JSPS KAKENHI under grant numbers 16K19429 and 18K15846. This work was also supported by funding from Mr. Hiroshi Mikitani, Mr. Marc Benioff, the L. K. Whittier Foundation, the Roddenberry Foundation, the Gladstone Institutes, the National Heart, Lung, and Blood Institute (NHLBI), and National Institutes of Health (NIH) (U01-HL100406, U01-HL098179, R01-HL130533, and R01-HL135358). Gladstone Institutes received support from National Center for Research Resources grant RR18928-01. A.K. designed and conceived this study, performed most of the experiments, and analyzed the data. R.H. and V.A. cultured cells and performed differentiation assays. M.O. performed and supported flow cytometry. T.M.K. and M.N. analyzed the RNA sequencing (RNA-seq), DNA methylation array, and SNP genotyping array data. J.K. H.H. and K.K. developed and provided the SeV vectors. Y.T. and K.T. provided and instructed experimental techniques. K.F. N.T. and S.Y. supervised the project. A.K. and S.Y. wrote the manuscript. A.K. and J.K. are co-inventors on a patent describing the method for producing naive human iPSCs from somatic cells. J.K. and H.H. are employees and K.K. is a board member of ID Pharma Co. Ltd. without compensation relating to this study. K.T. is on the scientific advisory board of I Peace, Inc. without salary. S.Y. is a scientific advisor to iPS Academia Japan without salary. We worked to ensure diversity in experimental samples through the selection of the cell lines.
Publisher Copyright:
© 2022 The Authors
PY - 2022/11/21
Y1 - 2022/11/21
N2 - Naive human induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with Sendai virus (SeV) vectors. However, only dermal fibroblasts have been successfully reprogrammed this way, and the process requires culture on feeder cells. Moreover, SeV vectors are highly persistent and inhibit subsequent differentiation of iPSCs. Here, we report a modified SeV vector system to generate transgene-free naive human iPSCs with superior differentiation potential. The modified method can be applied not only to fibroblasts but also to other somatic cell types. SeV vectors disappear quickly at early passages, and this approach enables the generation of naive iPSCs in a feeder-free culture. The naive iPSCs generated by this method show better differentiation to trilineage and extra-embryonic trophectoderm than those derived by conventional methods. This method can expand the application of iPSCs to research on early human development and regenerative medicine.
AB - Naive human induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with Sendai virus (SeV) vectors. However, only dermal fibroblasts have been successfully reprogrammed this way, and the process requires culture on feeder cells. Moreover, SeV vectors are highly persistent and inhibit subsequent differentiation of iPSCs. Here, we report a modified SeV vector system to generate transgene-free naive human iPSCs with superior differentiation potential. The modified method can be applied not only to fibroblasts but also to other somatic cell types. SeV vectors disappear quickly at early passages, and this approach enables the generation of naive iPSCs in a feeder-free culture. The naive iPSCs generated by this method show better differentiation to trilineage and extra-embryonic trophectoderm than those derived by conventional methods. This method can expand the application of iPSCs to research on early human development and regenerative medicine.
KW - LMYC
KW - Sendai virus vector
KW - extra-embryonic trophectoderm
KW - feeder-free culture
KW - hsa-microRNA-367
KW - induced pluripotent stem cells
KW - naive pluripotency
KW - reprogramming
KW - residual transgenes
KW - temperature sensitivity
UR - http://www.scopus.com/inward/record.url?scp=85142197199&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85142197199&partnerID=8YFLogxK
U2 - 10.1016/j.crmeth.2022.100317
DO - 10.1016/j.crmeth.2022.100317
M3 - Article
AN - SCOPUS:85142197199
SN - 2667-2375
VL - 2
JO - Cell Reports Methods
JF - Cell Reports Methods
IS - 11
M1 - 100317
ER -
