Robust detection of undifferentiated iPSC among differentiated cells

Sekine, Keisuke; Tsuzuki, Syusaku; Yasui, Ryota; Kobayashi, Tatsuya; Ikeda, Kazuki; Hamada, Yuki; Kanai, Eriko; Camp, J. Gray; Treutlein, Barbara; Ueno, Yasuharu; Okamoto, Satoshi; Taniguchi, Hideki

doi:10.1038/s41598-020-66845-6

“…We also demonstrated that the function of ESRG is not required for recapturing pluripotency via somatic cell reprogramming. Finally, the tightly regulated and high expression of ESRG promises to make an excellent marker of undifferentiated human PSCs both in basic research and clinical application [ 20 , 47 ].…”

Section: Discussionmentioning

confidence: 99%

The pluripotent stem cell-specific transcript ESRG is dispensable for human pluripotency

Takahashi

¹

,

Nakamura

²

,

Okubo

³

et al. 2021

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Human pluripotent stem cells (PSCs) express human endogenous retrovirus type-H (HERV-H), which exists as more than a thousand copies on the human genome and frequently produces chimeric transcripts as long-non-coding RNAs (lncRNAs) fused with downstream neighbor genes. Previous studies showed that HERV-H expression is required for the maintenance of PSC identity, and aberrant HERV-H expression attenuates neural differentiation potentials, however, little is known about the actual of function of HERV-H. In this study, we focused on ESRG, which is known as a PSC-related HERV-H-driven lncRNA. The global transcriptome data of various tissues and cell lines and quantitative expression analysis of PSCs showed that ESRG expression is much higher than other HERV-Hs and tightly silenced after differentiation. However, the loss of function by the complete excision of the entire ESRG gene body using a CRISPR/Cas9 platform revealed that ESRG is dispensable for the maintenance of the primed and naïve pluripotent states. The loss of ESRG hardly affected the global gene expression of PSCs or the differentiation potential toward trilineage. Differentiated cells derived from ESRG-deficient PSCs retained the potential to be reprogrammed into induced PSCs (iPSCs) by the forced expression of OCT3/4, SOX2, and KLF4. In conclusion, ESRG is dispensable for the maintenance and recapturing of human pluripotency.

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“…We also demonstrated that the function of ESRG is not required for recapturing pluripotency via somatic cell reprogramming. Finally, the tightly regulated and high expression of ESRG promises to make an excellent marker of undifferentiated PSCs both in basic research and clinical application [20, 38].…”

Section: Discussionmentioning

confidence: 99%

The pluripotent stem cell-specific transcript ESRG is dispensable for human pluripotency

Takahashi

¹

,

Nakamura

²

,

Narita

³

et al. 2020

Preprint

4

7

0

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Human pluripotent stem cells (PSCs) express human endogenous retrovirus type-H (HERV-H), which exists as more than a thousand copies on the human genome and frequently produces chimeric transcripts as long-non-coding RNAs (lncRNAs) fused with downstream neighbor genes. Previous studies showed that HERV-H expression is required for the maintenance of PSC identity, and aberrant HERV-H expression attenuates neural differentiation potentials, however, little is known about the actual of function of HERV-H. In this study, we focused on ESRG, which is known as a PSC-related HERV-H-driven lncRNA. The global transcriptome data of various tissues and cell lines and quantitative expression analysis of PSCs showed that ESRG expression is much higher than other HERV-Hs and tightly silenced after differentiation. However, the loss of function by the complete excision of the entire ESRG gene body using a CRISPR/Cas9 platform revealed that ESRG is dispensable for the maintenance of the primed and naïve pluripotent states. The loss of ESRG hardly affected the global gene expression of PSCs or the differentiation potential toward trilineage. Differentiated cells derived from ESRG-deficient PSCs retained the potential to be reprogrammed into induced PSCs (iPSCs) by the forced expression of OCT3/4, SOX2, and KLF4. In conclusion, ESRG is dispensable for the maintenance and recapturing of human pluripotency.

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“…To mitigate the potential risk of genomic instability and accumulation of oncogenic mutations [ 65 , 79 ], any iPSC therapy must, however, be extensively genetically screened prior to use [ 48 , 50 ] and any undifferentiated cells need to be detected by robust methods [ 80 ].…”

Section: Alternative Sources Of Schwann Cellsmentioning

confidence: 99%

Perspective on Schwann Cells Derived from Induced Pluripotent Stem Cells in Peripheral Nerve Tissue Engineering

Huang

¹

,

Powell

²

,

Phillips

³

et al. 2020

Cells

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Schwann cells play a crucial role in successful peripheral nerve repair and regeneration by supporting both axonal growth and myelination. Schwann cells are therefore a feasible option for cell therapy treatment of peripheral nerve injury. However, sourcing human Schwann cells at quantities required for development beyond research is challenging. Due to their availability, rapid in vitro expansion, survival, and integration within the host tissue, stem cells have attracted considerable attention as candidate cell therapies. Among them, induced pluripotent stem cells (iPSCs) with the associated prospects for personalized treatment are a promising therapy to take the leap from bench to bedside. In this critical review, we firstly focus on the current knowledge of the Schwann cell phenotype in regard to peripheral nerve injury, including crosstalk with the immune system during peripheral nerve regeneration. Then, we review iPSC to Schwann cell derivation protocols and the results from recent in vitro and in vivo studies. We finally conclude with some prospects for the use of iPSCs in clinical settings.

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Robust detection of undifferentiated iPSC among differentiated cells

Cited by 45 publications

References 31 publications

The pluripotent stem cell-specific transcript ESRG is dispensable for human pluripotency

The pluripotent stem cell-specific transcript ESRG is dispensable for human pluripotency

The pluripotent stem cell-specific transcript ESRG is dispensable for human pluripotency

Perspective on Schwann Cells Derived from Induced Pluripotent Stem Cells in Peripheral Nerve Tissue Engineering

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