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Junlin Yang
Hangzhou Normal University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-3421-7166
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Background: Reprogramming somatic cells to induced pluripotent stem cells (iPSCs) has opened new therapeutic possibilities. However, karyotypic abnormalities detected in iPSCs compromised their utility, especially chromosomal aberrations found at early passages raised serious safety concerns. The mechanism underlying the chromosomal abnormality in early-passage iPSCs is not known.
Methods: Human dermal fibroblasts (HDFs) were stimulated with KMOS (KLF4, cMYC, OCT4 and SOX2) proteins to enhance their proliferative capacity and many vigorous clones were obtained. Clonal reprogramming was carried out by KMOS mRNAs transfection to confirm the ‘chromosomal mutagenicity’ of reprogramming process. Subculturing was performed to examine karyotypic stability of iPSCs after the re-establishment of stemness. And antioxidant N-acetyl-cysteine (NAC) was added to the culture medium for further confirmming the mutagenicity in the first few days of reprogramming.
Results: Chromosomal aberrations were found in a small percentage of newly induced iPS clones by reprogramming transcription factors. Clonal reprogramming ruled out the aberrant chromosomes inherited from rare karyotypically abnormal parental cell subpopulation. More importantly, the antioxidant NAC effectively reduced the occurrence of chromosomal aberrations at the early stage of reprogramming. Once iPS cell lines were established, they restored karyotypic stability in subsequent subculturing.
Conclusions: Our results provided the first line of evidence for the ‘chromosomal mutagenicity’ of reprogramming process.
Keywords
induced pluripotent stem cell, reprogramme, karyotype, chromosomal aberration, genetic stability
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CURRENT STATUS: Published
View the published article at Cell Division.
Version 2
Posted 26 Oct, 2020
No community comments so far
Editorial decision: Accept
On 20 Oct, 2020
Editor assigned
On 16 Oct, 2020
Submission checks complete
On 15 Oct, 2020
Editor invited
On 15 Oct, 2020
No comments provided
Editorial decision: Major revision
On 14 Jul, 2020
Review #2 received
Received 09 Jul, 2020
Review #3 received
Received 09 Jul, 2020
Review #4 received
Received 04 Jul, 2020
Reviewer #4 agreed
On 27 Jun, 2020
Reviewer #2 agreed
On 24 Jun, 2020
Reviewer #3 agreed
On 24 Jun, 2020
Reviewers invited
Invitations sent on 03 Jun, 2020
Reviewer #1 agreed
On 03 Jun, 2020
Review #1 received
Received 03 Jun, 2020
Editor assigned
On 02 Jun, 2020
First submitted
On 01 Jun, 2020
Submission checks complete
On 01 Jun, 2020
Editor invited
On 01 Jun, 2020
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Subject Areas
Stem Cell & Developmental Cell Biology
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A new preprint design is coming!
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See the published version of this preprint at Cell Division.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Junlin Yang
Hangzhou Normal University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-3421-7166
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Cell Division.
Version 2
Posted 26 Oct, 2020
No community comments so far
Editorial decision: Accept
On 20 Oct, 2020
Editor assigned
On 16 Oct, 2020
Submission checks complete
On 15 Oct, 2020
Editor invited
On 15 Oct, 2020
No comments provided
Editorial decision: Major revision
On 14 Jul, 2020
Review #2 received
Received 09 Jul, 2020
Review #3 received
Received 09 Jul, 2020
Review #4 received
Received 04 Jul, 2020
Reviewer #4 agreed
On 27 Jun, 2020
Reviewer #2 agreed
On 24 Jun, 2020
Reviewer #3 agreed
On 24 Jun, 2020
Reviewers invited
Invitations sent on 03 Jun, 2020
Reviewer #1 agreed
On 03 Jun, 2020
Review #1 received
Received 03 Jun, 2020
Editor assigned
On 02 Jun, 2020
First submitted
On 01 Jun, 2020
Submission checks complete
On 01 Jun, 2020
Editor invited
On 01 Jun, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Stem Cell & Developmental Cell Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Cell Division.
Background: Reprogramming somatic cells to induced pluripotent stem cells (iPSCs) has opened new therapeutic possibilities. However, karyotypic abnormalities detected in iPSCs compromised their utility, especially chromosomal aberrations found at early passages raised serious safety concerns. The mechanism underlying the chromosomal abnormality in early-passage iPSCs is not known.
Methods: Human dermal fibroblasts (HDFs) were stimulated with KMOS (KLF4, cMYC, OCT4 and SOX2) proteins to enhance their proliferative capacity and many vigorous clones were obtained. Clonal reprogramming was carried out by KMOS mRNAs transfection to confirm the ‘chromosomal mutagenicity’ of reprogramming process. Subculturing was performed to examine karyotypic stability of iPSCs after the re-establishment of stemness. And antioxidant N-acetyl-cysteine (NAC) was added to the culture medium for further confirmming the mutagenicity in the first few days of reprogramming.
Results: Chromosomal aberrations were found in a small percentage of newly induced iPS clones by reprogramming transcription factors. Clonal reprogramming ruled out the aberrant chromosomes inherited from rare karyotypically abnormal parental cell subpopulation. More importantly, the antioxidant NAC effectively reduced the occurrence of chromosomal aberrations at the early stage of reprogramming. Once iPS cell lines were established, they restored karyotypic stability in subsequent subculturing.
Conclusions: Our results provided the first line of evidence for the ‘chromosomal mutagenicity’ of reprogramming process.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
This is a list of supplementary files associated with this preprint. Click to download.
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