An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness

Torre, Asier Galarza; Shaw, Joshua E; Wood, A.K.; Gilbert, Hamish T. J.; Dobre, Oana; Genever, Paul G.; Brennan, Keith; Richardson, Stephen M.; Swift, Joe

doi:10.1038/s41598-018-27346-9

Cited by 36 publications

(27 citation statements)

References 53 publications

(88 reference statements)

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“…We also investigated the effects of SUN2 overexpression (OE), using immortalized hMSCs (Y201 line) that maintain the multipotency and mechano-responsiveness of primary MSCs 49,50 . The proteomes of Y201 cells were quantified with doxycycline-induced OE of SUN2 to 160% and 410% of control levels (Fig.…”

Section: Resultsmentioning

confidence: 99%

Nuclear decoupling is part of a rapid protein-level cellular response to high-intensity mechanical loading

et al. 2019

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Studies of cellular mechano-signaling have often utilized static models that do not fully replicate the dynamics of living tissues. Here, we examine the time-dependent response of primary human mesenchymal stem cells (hMSCs) to cyclic tensile strain (CTS). At low-intensity strain (1 h, 4% CTS at 1 Hz), cell characteristics mimic responses to increased substrate stiffness. As the strain regime is intensified (frequency increased to 5 Hz), we characterize rapid establishment of a broad, structured and reversible protein-level response, even as transcription is apparently downregulated. Protein abundance is quantified coincident with changes to protein conformation and post-translational modification (PTM). Furthermore, we characterize changes to the linker of nucleoskeleton and cytoskeleton (LINC) complex that bridges the nuclear envelope, and specifically to levels and PTMs of Sad1/UNC-84 (SUN) domain-containing protein 2 (SUN2). The result of this regulation is to decouple mechano-transmission between the cytoskeleton and the nucleus, thus conferring protection to chromatin.

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Section: Resultsmentioning

confidence: 99%

Nuclear decoupling is part of a rapid protein-level cellular response to high-intensity mechanical loading

et al. 2019

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“…The osteogenic potential of immortalized MSCs is usually equal or higher than that of primary MSCs [ 16 , 54 ], while their chondrogenic potential is generally poor [ 17 , 21 , 26 , 55 ]. Immortalized MSCs and osteoprogenitor cells have often been reported to be able to differentiate into adipocytes [ 49 , 56 , 57 , 58 , 59 ], but adipogenic potential may be reduced after immortalization [ 26 , 60 , 61 ].…”

Section: Discussionmentioning

confidence: 99%

Generation of Mesenchymal Cell Lines Derived from Aged Donors

Piñeiro-Ramil

Sanjurjo‐Rodríguez

Rodríguez-Fernández

et al. 2021

IJMS

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Background: Mesenchymal stromal cells (MSCs) have the capacity for self-renewal and multi-differentiation, and for this reason they are considered a potential cellular source in regenerative medicine of cartilage and bone. However, research on this field is impaired by the predisposition of primary MSCs to senescence during culture expansion. Therefore, the aim of this study was to generate and characterize immortalized MSC (iMSC) lines from aged donors. Methods: Primary MSCs were immortalized by transduction of simian virus 40 large T antigen (SV40LT) and human telomerase reverse transcriptase (hTERT). Proliferation, senescence, phenotype and multi-differentiation potential of the resulting iMSC lines were analyzed. Results: MSCs proliferate faster than primary MSCs, overcome senescence and are phenotypically similar to primary MSCs. Nevertheless, their multi-differentiation potential is unbalanced towards the osteogenic lineage. There are no clear differences between osteoarthritis (OA) and non-OA iMSCs in terms of proliferation, senescence, phenotype or differentiation potential. Conclusions: Primary MSCs obtained from elderly patients can be immortalized by transduction of SV40LT and hTERT. The high osteogenic potential of iMSCs converts them into an excellent cellular source to take part in in vitro models to study bone tissue engineering.

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“…Positive for ARS [19], ALP activity [19,82,83], and Runx2 upregulation [19,82] Positive for ABS, GAG assay, and Sox9 upregulation in pellet culture [19] Positive for OROS (reduced compared to primary MSCs) and upregulation of adipogenesis-related genes [19,82] Y101 (derived from the same donor than Y201)…”

Section: Y201mentioning

confidence: 99%

Usefulness of Mesenchymal Cell Lines for Bone and Cartilage Regeneration Research

Piñeiro-Ramil

Sanjurjo‐Rodríguez

Castro-Viñuelas

et al. 2019

IJMS

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The unavailability of sufficient numbers of human primary cells is a major roadblock for in vitro repair of bone and/or cartilage, and for performing disease modelling experiments. Immortalized mesenchymal stromal cells (iMSCs) may be employed as a research tool for avoiding these problems. The purpose of this review was to revise the available literature on the characteristics of the iMSC lines, paying special attention to the maintenance of the phenotype of the primary cells from which they were derived, and whether they are effectively useful for in vitro disease modeling and cell therapy purposes. This review was performed by searching on Web of Science, Scopus, and PubMed databases from 1 January 2015 to 30 September 2019. The keywords used were ALL = (mesenchymal AND ("cell line" OR immortal*) AND (cartilage OR chondrogenesis OR bone OR osteogenesis) AND human). Only original research studies in which a human iMSC line was employed for osteogenesis or chondrogenesis experiments were included. After describing the success of the immortalization protocol, we focused on the iMSCs maintenance of the parental phenotype and multipotency. According to the literature revised, it seems that the maintenance of these characteristics is not guaranteed by immortalization, and that careful selection and validation of clones with particular characteristics is necessary for taking advantage of the full potential of iMSC to be employed in bone and cartilage-related research.

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An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness

Cited by 36 publications

References 53 publications

Nuclear decoupling is part of a rapid protein-level cellular response to high-intensity mechanical loading

Nuclear decoupling is part of a rapid protein-level cellular response to high-intensity mechanical loading

Generation of Mesenchymal Cell Lines Derived from Aged Donors

Usefulness of Mesenchymal Cell Lines for Bone and Cartilage Regeneration Research

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