Generation of transgene-free porcine intermediate type induced pluripotent stem cells

Research output: Contribution to journalJournal article

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Generation of transgene-free porcine intermediate type induced pluripotent stem cells. / Li, Dong; Secher, Jan; Hyttel, Poul; Ivask, Marilin; Kolko, Miriam; Hall, Vanessa Jane; Freude, Kristine K.

In: Cell Cycle, Vol. 17, No. 23, 2018, p. 2547-2563.

Research output: Contribution to journalJournal article

Harvard

Li, D, Secher, J, Hyttel, P, Ivask, M, Kolko, M, Hall, VJ & Freude, KK 2018, 'Generation of transgene-free porcine intermediate type induced pluripotent stem cells', Cell Cycle, vol. 17, no. 23, pp. 2547-2563. https://doi.org/10.1080/15384101.2018.1548790

APA

Li, D., Secher, J., Hyttel, P., Ivask, M., Kolko, M., Hall, V. J., & Freude, K. K. (2018). Generation of transgene-free porcine intermediate type induced pluripotent stem cells. Cell Cycle, 17(23), 2547-2563. https://doi.org/10.1080/15384101.2018.1548790

Vancouver

Li D, Secher J, Hyttel P, Ivask M, Kolko M, Hall VJ et al. Generation of transgene-free porcine intermediate type induced pluripotent stem cells. Cell Cycle. 2018;17(23):2547-2563. https://doi.org/10.1080/15384101.2018.1548790

Author

Li, Dong ; Secher, Jan ; Hyttel, Poul ; Ivask, Marilin ; Kolko, Miriam ; Hall, Vanessa Jane ; Freude, Kristine K. / Generation of transgene-free porcine intermediate type induced pluripotent stem cells. In: Cell Cycle. 2018 ; Vol. 17, No. 23. pp. 2547-2563.

Bibtex

@article{9301639434e140a39c029cbc015ba896,
title = "Generation of transgene-free porcine intermediate type induced pluripotent stem cells",
abstract = "Physiologically and anatomically, humans and pigs share many similarities, which make porcine induced pluripotent stem cells (piPSCs) very attractive for modeling human cell therapy as well as for testing safety of iPSC based cell replacement therapies. To date, several integrative and non-integrative strategies have been reported to successfully generate piPSCs, but all resulting piPSCs had integration of transgenes. The use of integrative methods has the disadvantage of potential lack of silencing or inappropriate re-activation of these genes during differentiation, as well as uncertainty regarding disruption of important genomic regions caused by integration. In our study, we performed a non-integrative vector based reprogramming approach using porcine fetal fibroblasts. The resulting four piPSC lines were positive for pluripotency marker and when subjected to in vitro and in vivo differentiation assays, all four lines formed embryoid bodies, capable to differentiate into all three germ layers, and three out of the four cell lines formed teratomas. PCR analysis on genomic and plasmid DNA revealed that the episomal vectors were undetectable in six out of eight subclones derived from one of the piPSC lines (piPSC1) above passage 20. These piPSCs could potentially be ideal cell lines for the generation of porcine in vitro and in vivo models. Furthermore, subsequent analyses of our new transgene independent piPSCs could provide novel insights on the genetic and epigenetic necessities to achieve and maintain piPSCs.",
keywords = "in vitro differentiation, non-integrative strategies, Porcine induced pluripotent stem cells, transgene-free piPSCs",
author = "Dong Li and Jan Secher and Poul Hyttel and Marilin Ivask and Miriam Kolko and Hall, {Vanessa Jane} and Freude, {Kristine K.}",
year = "2018",
doi = "10.1080/15384101.2018.1548790",
language = "English",
volume = "17",
pages = "2547--2563",
journal = "Cell Cycle",
issn = "1538-4101",
publisher = "Taylor & Francis",
number = "23",

}

RIS

TY - JOUR

T1 - Generation of transgene-free porcine intermediate type induced pluripotent stem cells

AU - Li, Dong

AU - Secher, Jan

AU - Hyttel, Poul

AU - Ivask, Marilin

AU - Kolko, Miriam

AU - Hall, Vanessa Jane

AU - Freude, Kristine K.

PY - 2018

Y1 - 2018

N2 - Physiologically and anatomically, humans and pigs share many similarities, which make porcine induced pluripotent stem cells (piPSCs) very attractive for modeling human cell therapy as well as for testing safety of iPSC based cell replacement therapies. To date, several integrative and non-integrative strategies have been reported to successfully generate piPSCs, but all resulting piPSCs had integration of transgenes. The use of integrative methods has the disadvantage of potential lack of silencing or inappropriate re-activation of these genes during differentiation, as well as uncertainty regarding disruption of important genomic regions caused by integration. In our study, we performed a non-integrative vector based reprogramming approach using porcine fetal fibroblasts. The resulting four piPSC lines were positive for pluripotency marker and when subjected to in vitro and in vivo differentiation assays, all four lines formed embryoid bodies, capable to differentiate into all three germ layers, and three out of the four cell lines formed teratomas. PCR analysis on genomic and plasmid DNA revealed that the episomal vectors were undetectable in six out of eight subclones derived from one of the piPSC lines (piPSC1) above passage 20. These piPSCs could potentially be ideal cell lines for the generation of porcine in vitro and in vivo models. Furthermore, subsequent analyses of our new transgene independent piPSCs could provide novel insights on the genetic and epigenetic necessities to achieve and maintain piPSCs.

AB - Physiologically and anatomically, humans and pigs share many similarities, which make porcine induced pluripotent stem cells (piPSCs) very attractive for modeling human cell therapy as well as for testing safety of iPSC based cell replacement therapies. To date, several integrative and non-integrative strategies have been reported to successfully generate piPSCs, but all resulting piPSCs had integration of transgenes. The use of integrative methods has the disadvantage of potential lack of silencing or inappropriate re-activation of these genes during differentiation, as well as uncertainty regarding disruption of important genomic regions caused by integration. In our study, we performed a non-integrative vector based reprogramming approach using porcine fetal fibroblasts. The resulting four piPSC lines were positive for pluripotency marker and when subjected to in vitro and in vivo differentiation assays, all four lines formed embryoid bodies, capable to differentiate into all three germ layers, and three out of the four cell lines formed teratomas. PCR analysis on genomic and plasmid DNA revealed that the episomal vectors were undetectable in six out of eight subclones derived from one of the piPSC lines (piPSC1) above passage 20. These piPSCs could potentially be ideal cell lines for the generation of porcine in vitro and in vivo models. Furthermore, subsequent analyses of our new transgene independent piPSCs could provide novel insights on the genetic and epigenetic necessities to achieve and maintain piPSCs.

KW - in vitro differentiation

KW - non-integrative strategies

KW - Porcine induced pluripotent stem cells

KW - transgene-free piPSCs

U2 - 10.1080/15384101.2018.1548790

DO - 10.1080/15384101.2018.1548790

M3 - Journal article

C2 - 30457474

AN - SCOPUS:85058109545

VL - 17

SP - 2547

EP - 2563

JO - Cell Cycle

JF - Cell Cycle

SN - 1538-4101

IS - 23

ER -

ID: 210377413