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Immortalization of mesenchymal stromal cells by hTERT does not affect the functional properties of secreted extracellular vesicles.

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Title: Immortalization of mesenchymal stromal cells by hTERT does not affect the functional properties of secreted extracellular vesicles.
Authors: Brancolini, Alessia1 (AUTHOR), Bobbili, Madhusudhan Reddy2,3,4 (AUTHOR), Pultar, Marianne5 (AUTHOR), Mazidi, Zahra1 (AUTHOR), Wieser, Matthias1 (AUTHOR), Gamauf, Johanna1 (AUTHOR), Roefs, Marieke Theodora1 (AUTHOR), Corso, Giulia1 (AUTHOR), Nivarthi, Harini1 (AUTHOR), Paredes, Maria Belen Arteaga4,6 (AUTHOR), Bettelheim, Dieter7 (AUTHOR), Arcalis, Elsa8 (AUTHOR), Dmitry, Sivun9 (AUTHOR), Jacak, Jaroslaw3,9 (AUTHOR), Gerner, Iris4,6 (AUTHOR), Bileck, Andrea10 (AUTHOR), Gerner, Christoper10 (AUTHOR), Hackl, Matthias4,5 (AUTHOR), Grillari, Johannes1,2,3,4 (AUTHOR) johannes.grillari@lbg.ac.at, Grillari-Voglauer, Regina1,4 (AUTHOR) regina.grillari@evercyte.com
Source: Journal of Biotechnology. Jun2026, Vol. 414, p124-145. 22p.
Subjects: Extracellular vesicles, Telomerase reverse transcriptase, Cell physiology, Cell proliferation, Cellular therapy, Therapeutics, Mesenchymal stem cells
Abstract: Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) have emerged as promising and safe therapeutic agents, however, donor heterogeneities, limited replicative life span and changes in the cellular phenotype throughout in vitro cultivation remain major hurdles for scalable EV production. For these reasons, this study aims to investigate the use of hTERT immortalized ('telomerized') MSCs as a potential source for efficient, standardized, reliable MSC-EVs production by comparing parental primary to their telomerized MSC counterparts. We observed that hTERT expression does not affect cell morphology or cellular doubling time, while ensuring unlimited, stable in vitro propagation. In addition, telomerized WJ-MSCs maintained the canonical expression profile of surface markers and the tri-lineage differentiation potential of their primary counterparts. In terms of EV characteristics, the immortalization by hTERT expression did not affect size, number, cargo composition or biological activity regarding anti-inflammatory, anti-fibrotic and wound healing properties in vitro. In summary, the use of hTERT to immortalize MSCs leads to the creation of cell lines that continuously produce MSC-EVs without altering any key functionalities of the cells or resulting EVs. This suggests that telomerization of human cells from single donors is a promising strategy for generating cell factories that can produce EVs in standardized conditions at scale and with standardization. • hTERT immortalization alone ensures stable, unlimited in vitro propagation of MSCs • MSC-markers and differentiation are maintained upon hTERT immortalization • Immortalization by hTERT expression do not affect size, number or miRNA content of EVs • Biological activity of MSC-EVs is not affected by hTERT immortalization • Safety profile of MSCs and EVs is not affected by hTERT immortalization [ABSTRACT FROM AUTHOR]
Copyright of Journal of Biotechnology is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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DbLabel: Engineering Source
An: 193007092
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PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
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  Label: Title
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  Data: Immortalization of mesenchymal stromal cells by hTERT does not affect the functional properties of secreted extracellular vesicles.
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  Data: <searchLink fieldCode="AR" term="%22Brancolini%2C+Alessia%22">Brancolini, Alessia</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Bobbili%2C+Madhusudhan+Reddy%22">Bobbili, Madhusudhan Reddy</searchLink><relatesTo>2,3,4</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Pultar%2C+Marianne%22">Pultar, Marianne</searchLink><relatesTo>5</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Mazidi%2C+Zahra%22">Mazidi, Zahra</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wieser%2C+Matthias%22">Wieser, Matthias</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Gamauf%2C+Johanna%22">Gamauf, Johanna</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Roefs%2C+Marieke+Theodora%22">Roefs, Marieke Theodora</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Corso%2C+Giulia%22">Corso, Giulia</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Nivarthi%2C+Harini%22">Nivarthi, Harini</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Paredes%2C+Maria+Belen+Arteaga%22">Paredes, Maria Belen Arteaga</searchLink><relatesTo>4,6</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Bettelheim%2C+Dieter%22">Bettelheim, Dieter</searchLink><relatesTo>7</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Arcalis%2C+Elsa%22">Arcalis, Elsa</searchLink><relatesTo>8</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Dmitry%2C+Sivun%22">Dmitry, Sivun</searchLink><relatesTo>9</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Jacak%2C+Jaroslaw%22">Jacak, Jaroslaw</searchLink><relatesTo>3,9</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Gerner%2C+Iris%22">Gerner, Iris</searchLink><relatesTo>4,6</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Bileck%2C+Andrea%22">Bileck, Andrea</searchLink><relatesTo>10</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Gerner%2C+Christoper%22">Gerner, Christoper</searchLink><relatesTo>10</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Hackl%2C+Matthias%22">Hackl, Matthias</searchLink><relatesTo>4,5</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Grillari%2C+Johannes%22">Grillari, Johannes</searchLink><relatesTo>1,2,3,4</relatesTo> (AUTHOR)<i> johannes.grillari@lbg.ac.at</i><br /><searchLink fieldCode="AR" term="%22Grillari-Voglauer%2C+Regina%22">Grillari-Voglauer, Regina</searchLink><relatesTo>1,4</relatesTo> (AUTHOR)<i> regina.grillari@evercyte.com</i>
– Name: TitleSource
  Label: Source
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  Data: <searchLink fieldCode="JN" term="%22Journal+of+Biotechnology%22">Journal of Biotechnology</searchLink>. Jun2026, Vol. 414, p124-145. 22p.
– Name: Subject
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  Data: <searchLink fieldCode="DE" term="%22Extracellular+vesicles%22">Extracellular vesicles</searchLink><br /><searchLink fieldCode="DE" term="%22Telomerase+reverse+transcriptase%22">Telomerase reverse transcriptase</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+physiology%22">Cell physiology</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+proliferation%22">Cell proliferation</searchLink><br /><searchLink fieldCode="DE" term="%22Cellular+therapy%22">Cellular therapy</searchLink><br /><searchLink fieldCode="DE" term="%22Therapeutics%22">Therapeutics</searchLink><br /><searchLink fieldCode="DE" term="%22Mesenchymal+stem+cells%22">Mesenchymal stem cells</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) have emerged as promising and safe therapeutic agents, however, donor heterogeneities, limited replicative life span and changes in the cellular phenotype throughout in vitro cultivation remain major hurdles for scalable EV production. For these reasons, this study aims to investigate the use of hTERT immortalized ('telomerized') MSCs as a potential source for efficient, standardized, reliable MSC-EVs production by comparing parental primary to their telomerized MSC counterparts. We observed that hTERT expression does not affect cell morphology or cellular doubling time, while ensuring unlimited, stable in vitro propagation. In addition, telomerized WJ-MSCs maintained the canonical expression profile of surface markers and the tri-lineage differentiation potential of their primary counterparts. In terms of EV characteristics, the immortalization by hTERT expression did not affect size, number, cargo composition or biological activity regarding anti-inflammatory, anti-fibrotic and wound healing properties in vitro. In summary, the use of hTERT to immortalize MSCs leads to the creation of cell lines that continuously produce MSC-EVs without altering any key functionalities of the cells or resulting EVs. This suggests that telomerization of human cells from single donors is a promising strategy for generating cell factories that can produce EVs in standardized conditions at scale and with standardization. • hTERT immortalization alone ensures stable, unlimited in vitro propagation of MSCs • MSC-markers and differentiation are maintained upon hTERT immortalization • Immortalization by hTERT expression do not affect size, number or miRNA content of EVs • Biological activity of MSC-EVs is not affected by hTERT immortalization • Safety profile of MSCs and EVs is not affected by hTERT immortalization [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Journal of Biotechnology is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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      – Type: doi
        Value: 10.1016/j.jbiotec.2026.03.010
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      – Code: eng
        Text: English
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      Pagination:
        PageCount: 22
        StartPage: 124
    Subjects:
      – SubjectFull: Extracellular vesicles
        Type: general
      – SubjectFull: Telomerase reverse transcriptase
        Type: general
      – SubjectFull: Cell physiology
        Type: general
      – SubjectFull: Cell proliferation
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      – SubjectFull: Cellular therapy
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      – SubjectFull: Therapeutics
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      – SubjectFull: Mesenchymal stem cells
        Type: general
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      – TitleFull: Immortalization of mesenchymal stromal cells by hTERT does not affect the functional properties of secreted extracellular vesicles.
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              Text: Jun2026
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