Telomere Length Defines the Cardiomyocyte Differentiation Potency of Mouse Induced Pluripotent Stem Cells

Aguado, Tania; Gutiérrez, Francisco J.; Aix, Esther; Schneider, Ralph P.; Giovinazzo, Giovanna; Blasco, María A.; Flores, Ignacio

doi:10.1002/stem.2497

Cited by 19 publications

(17 citation statements)

References 71 publications

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“…In agreement, Zeng et al demonstrated that telomeres were elongated during in vitro expansion and were then stabilized through a telomerase-dependent mechanism [32]. Our 2D and 3D expansion and directed differentiation of the revived cells demonstrated stable expression of telomerase and telomere length, as several studies have reported a correlation between short telomeres and unstable directed differentiation in mouse and human pluripotent cells [33,34].…”

Section: Discussionsupporting

confidence: 90%

Long-Term Stability and Differentiation Potential of Cryopreserved cGMP-Compliant Human Induced Pluripotent Stem Cells

Shafa

Walsh

Panchalingam

et al. 2019

IJMS

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The clinical effectiveness of human induced pluripotent stem cells (iPSCs) is highly dependent on a few key quality characteristics including the generation of high quality cell bank, long-term genomic stability, post-thaw viability, plating efficiency, retention of pluripotency, directed differentiation, purity, potency, and sterility. We have already reported the establishment of iPSC master cell banks (MCBs) and working cell banks (WCBs) under current good manufacturing procedure (cGMP)-compliant conditions. In this study, we assessed the cellular and genomic stability of the iPSC lines generated and cryopreserved five years ago under cGMP-compliant conditions. iPSC lines were thawed, characterized, and directly differentiated into cells from three germ layers including cardiomyocytes (CMs), neural stem cells (NSCs), and definitive endoderm (DE). The cells were also expanded in 2D and 3D spinner flasks to evaluate their long-term expansion potential in matrix-dependent and feeder-free culture environment. All three lines successfully thawed and attached to the L7TM matrix, and formed typical iPSC colonies that expressed pluripotency markers over 15 passages. iPSCs maintained their differentiation potential as demonstrated with spontaneous and directed differentiation to the three germ layers and corresponding expression of specific markers, respectfully. Furthermore, post-thaw cells showed normal karyotype, negative mycoplasma, and sterility testing. These cells maintained both their 2D and 3D proliferation potential after five years of cryopreservation without acquiring karyotype abnormality, loss of pluripotency, and telomerase activity. These results illustrate the long-term stability of cGMP iPSC lines, which is an important step in establishing a reliable, long-term source of starting materials for clinical and commercial manufacturing of iPSC-derived cell therapy products.

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Section: Discussionsupporting

confidence: 90%

Long-Term Stability and Differentiation Potential of Cryopreserved cGMP-Compliant Human Induced Pluripotent Stem Cells

Shafa

Walsh

Panchalingam

et al. 2019

IJMS

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“…Similarly, Imaizumi et al [40] and López et al [41] both observed no chromosomal structural abnormalities in cryopreserved human induced pluripotent stem cells and adipose-derived stem cells, respectively. Recently, it was shown that the differentiation potency of induced pluripotent stem cells was correlated with their telomere length [42]. These findings, coupled with our experiment data, offer convincing evidence that our cPDLSC sheets could maintain similar differentiation capacity as fPDLSC sheets.…”

Section: Discussionsupporting

confidence: 85%

Effect of cryopreservation on proliferation and differentiation of periodontal ligament stem cell sheets

Li¹,

Feng²,

Gu³

et al. 2017

Stem Cell Res Ther

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BackgroundCryopreservation has been extensively applied to the long-term storage of a diverse range of biological materials. However, no comprehensive study is currently available on the cryopreservation of periodontal ligament stem cell (PDLSC) sheets which have been suggested as excellent transplant materials for periodontal tissue regeneration. The aim of this study is to investigate the effect of cryopreservation on the structural integrity and functional viability of PDLSC sheets.MethodsPDLSC sheets prepared from extracted human molars were divided into two groups: the cryopreservation group (cPDLSC sheets) and the freshly prepared control group (fPDLSC sheets). The cPDLSC sheets were cryopreserved in a solution consisting of 90% fetal bovine serum and 10% dimethyl sulfoxide for 3 months. Cell viability and cell proliferation rates of PDLSCs in both groups were evaluated by cell viability assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, respectively. The multilineage differentiation potentials of the cells were assessed by von Kossa staining and Oil Red O staining. The chromosomal stability was examined by karyotype analysis. Moreover, the cell sheets in each group were transplanted subcutaneously into the dorsal site of nude mice, after which Sirius Red staining was performed to analyze the efficiency of tissue regeneration.ResultsThe PDLSCs derived from both groups of cell sheets showed no significant difference in their viability, proliferative capacities, and multilineage differentiation potentials, as well as chromosomal stability. Furthermore, transplantation experiments based on a mouse model demonstrated that the cPDLSC sheets were equally effective in generating viable osteoid tissues in vivo as their freshly prepared counterparts. In both cases, the regenerated tissues showed similar network patterns of bone-like matrix.ConclusionsOur results offer convincing evidence that cryopreservation does not alter the biological properties of PDLSC sheets and could enhance their clinical utility in tissue regeneration.

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“…For example, Mosteiro and colleagues found that senescence promoted in vivo programming of murine iPSCs, although the relevance of telomere integrity in this process was not directly assessed (Mosteiro et al, 2018). In cardiomyocytes stimulated to undergo iPSC reprogramming, there was an inverse correlation between reprogramming potential and telomere length (Aguado et al, 2016). In primary human cells undergoing senescence, H3K27me3 loss is correlated with gene expression changes that may drive senescence, and depletion of EZH2 contributes to the Senescence-Associated Secretory Phenotype (SASP) (Ito et al, 2018;Shah et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Telomere dysfunction cooperates with epigenetic alterations to impair murine embryonic stem cell fate commitment

Criqui

Qamra

Chu

et al. 2020

eLife

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The precise relationship between epigenetic alterations and telomere dysfunction is still an extant question. Previously, we showed that eroded telomeres lead to differentiation instability in murine embryonic stem cells (mESCs) via DNA hypomethylation at pluripotency-factor promoters. Here, we uncovered that telomerase reverse transcriptase null (Tert-/-) mESCs exhibit genome-wide alterations in chromatin accessibility and gene expression during differentiation. These changes were accompanied by an increase of H3K27me3 globally, an altered chromatin landscape at the Pou5f1/Oct4 promoter, and a refractory response to differentiation cues. Inhibition of the Polycomb Repressive Complex 2 (PRC2), an H3K27 tri-methyltransferase, exacerbated the impairment in differentiation and pluripotency gene repression in Tert-/- mESCs but not wild-type mESCs, whereas inhibition of H3K27me3 demethylation led to a partial rescue of the Tert-/- phenotype. These data reveal a new interdependent relationship between H3K27me3 and telomere integrity in stem cell lineage commitment that may have implications in aging and cancer.

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Telomere Length Defines the Cardiomyocyte Differentiation Potency of Mouse Induced Pluripotent Stem Cells

Cited by 19 publications

References 71 publications

Long-Term Stability and Differentiation Potential of Cryopreserved cGMP-Compliant Human Induced Pluripotent Stem Cells

Long-Term Stability and Differentiation Potential of Cryopreserved cGMP-Compliant Human Induced Pluripotent Stem Cells

Effect of cryopreservation on proliferation and differentiation of periodontal ligament stem cell sheets

Telomere dysfunction cooperates with epigenetic alterations to impair murine embryonic stem cell fate commitment

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