Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

Zhai, Weichao; Yong, Derrick; El-Jawhari, Jehan J.; Cuthbert, Richard; McGonagle, Dennis; Naing, May Win; Jones, Elena

doi:10.1016/j.jcyt.2019.05.001

Cited by 29 publications

(24 citation statements)

References 162 publications

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“…The elevated expression of MDM2, which promotes p53 degradation, might explain the low levels of apoptosis in the stressed populations. No p16 (CDKN2A) upregulation is seen, in agreement with the previous findings (Dulić et al, 2000;Alekseenko et al, 2012;Muñoz-Espín et al, 2013;Zhai et al, 2019), suggesting this is not a requisite marker for senescence. Interestingly, S phase genes such as PCNA and POLH are down-regulated in the constant HS group, arrested at G1.…”

Section: Hs Leads To Decreased Proliferation Cell Cycle Arrest and supporting

confidence: 91%

Heat Shock Alters Mesenchymal Stem Cell Identity and Induces Premature Senescence

Shimoni

Goldstein

Ribarski-Chorev

et al. 2020

Front. Cell Dev. Biol.

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Heat stress can have a serious impact on the health of both humans and animals. A major question is how heat stress affects normal development and differentiation at both the cellular and the organism levels. Here we use an in vitro experimental system to address how heat shock treatment influences the properties of bovine mesenchymal stem cells (MSCs)-multipotent progenitor cells-which are found in most tissues. Because cattle are sensitive to harsh external temperatures, studying the effects of heat shock on MSCs provides a unique platform to address cellular stress in a physiologically relevant model organism. Following isolation and characterization of MSCs from the cow's umbilical cord, heat shock was induced either as a pulse (1 h) or continuously (3 days), and consequent effects on MSCs were characterized. Heat shock induced extensive phenotypic changes in MSCs and dramatically curtailed their capacity to proliferate and differentiate. These changes were associated with a partial arrest in the G1/S or G2/M checkpoints. Furthermore, MSCs lost their ability to resolve the inflammatory response of RAW macrophages in coculture. A possible explanation for this loss of function is the accumulation of reactive oxygen species and malfunction of the mitochondria in the treated cells. Heat shock treatments resulted in stressinduced premature senescence, affecting the MSCs' ability to proliferate properly for many cell passages to follow. Exposure to elevated external temperatures leads to mitochondrial damage and oxidative stress, which in turn conveys critical changes in the proliferation, differentiation, and immunomodulatory phenotype of heat-stressed MSCs. A better understanding of the effect of heat shock on humans and animals may result in important health and economic benefits.

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Section: Hs Leads To Decreased Proliferation Cell Cycle Arrest and supporting

confidence: 91%

Heat Shock Alters Mesenchymal Stem Cell Identity and Induces Premature Senescence

Shimoni

Goldstein

Ribarski-Chorev

et al. 2020

Front. Cell Dev. Biol.

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“…These results indicated an age-related decline in MSC proliferation, confirming our colony number and integrated density data. A further study investigating the proportions of senescent cells in rare sorted CD45 low CD271 + cells from young and old donors, using emerging methodologies [58], will help to clarify this point.…”

Section: Discussionmentioning

confidence: 99%

The Analysis of In Vivo Aging in Human Bone Marrow Mesenchymal Stromal Cells Using Colony-Forming Unit-Fibroblast Assay and the CD45^lowCD271⁺ Phenotype

Ganguly

El-Jawhari

Burska

et al. 2019

Stem Cells International

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Uncultured mesenchymal stromal cells (MSCs) are increasingly used in therapies; however, the effects of donor age on their biological characteristics and gene expression remain unclear. The aim of this study was to investigate age-related changes in bone marrow (BM) MSCs following minimal or no culture manipulation. Iliac crest BM was aspirated from 67 healthy donors (19-89 years old) and directly used for the colony-forming unit-fibroblast (CFU-F) assay or CD45lowCD271+ cell enumeration. The colonies were analysed for colony area and integrated density (ID) when grown in standard MSC media or media supplemented with human serum from young (YS) or old (OS) donors. There was a notable age-related decline in the number of MSCs per millilitre of BM aspirate revealed by the CFU-F assay (r=−0.527, p<0.0001) or flow cytometry (r=−0.307, p=0.0116). Compared to young donors (19-40 years old), colony IDs were significantly lower in older donors (61-89 years old), particularly for smaller-sized colonies (42% lower, p<0.01). When cultured in media supplemented with OS, young and old donor MSCs formed colonies with lower IDs, by 21%, p<0.0001, and 27%, p<0.05, respectively, indicating the formation of smaller sparser colonies. No significant differences in the expression of selected adipogenic, osteogenic, stromal, and bone remodelling genes as well as CD295, CD146, CD106, and connexin 43 surface molecules were found in sorted CD45lowCD271+ MSCs from young and old donors (n=8 donors each). Altogether, these results show similar trends for age-related decline in BM MSC numbers measured by the CFU-F assay and flow cytometry and reveal age-related effects of human serum on MSC colony formation. No significant differences in selected gene expression in uncultured CD45lowCD271+ MSCs suggest that old donor MSCs may not be inferior in regard to their multipotential functions. Due to large donor-to-donor variation in all donor groups, our data indicate that an individual’s chronological age is not a reliable predictor of their MSC number or potency.

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“…Recently, novel specific biomarkers were found to demonstrate the senescent state of MSCs[ 21 ]. MSC-derived microvesicles (MVs) are one such biomarker[ 44 ], and senescent late passage MSCs displayed a smaller MSC-MV size compared with early passage MSCs.…”

Section: Characteristics and Functional Changes Of Aged Mscsmentioning

confidence: 99%

“…Additionally, MSCs from young donors may also become senescent because of excessive cell passage, oxidative stress, and other injuries[ 19 , 20 ]. The senescent cells manifest a sequence of progressive changes in cellular morphology, biological function, and molecular expression[ 21 , 22 ], as well as weakened efficacy in cell-based therapies[ 23 ]. Therefore, appropriate quality controls or cellular rejuvenation processes are required to obtain clinical-grade MSCs.…”

Section: Introductionmentioning

confidence: 99%

Senescent mesenchymal stem/stromal cells and restoring their cellular functions

Meng¹,

Liu²,

Lu³

et al. 2020

WJSC

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Mesenchymal stem/stromal cells (MSCs) have various properties that make them promising candidates for stem cell-based therapies in clinical settings. These include self-renewal, multilineage differentiation, and immunoregulation. However, recent studies have confirmed that aging is a vital factor that limits their function and therapeutic properties as standardized clinical products. Understanding the features of senescence and exploration of cell rejuvenation methods are necessary to develop effective strategies that can overcome the shortage and instability of MSCs. This review will summarize the current knowledge on characteristics and functional changes of aged MSCs. Additionally, it will highlight cell rejuvenation strategies such as molecular regulation, non-coding RNA modifications, and microenvironment controls that may enhance the therapeutic potential of MSCs in clinical settings.

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Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

Abstract: This is a repository copy of Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions.

Cited by 29 publications

References 162 publications

Heat Shock Alters Mesenchymal Stem Cell Identity and Induces Premature Senescence

Heat Shock Alters Mesenchymal Stem Cell Identity and Induces Premature Senescence

The Analysis of In Vivo Aging in Human Bone Marrow Mesenchymal Stromal Cells Using Colony-Forming Unit-Fibroblast Assay and the CD45^lowCD271⁺ Phenotype

Senescent mesenchymal stem/stromal cells and restoring their cellular functions

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Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

Abstract: This is a repository copy of Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions.

Cited by 29 publications

References 162 publications

Heat Shock Alters Mesenchymal Stem Cell Identity and Induces Premature Senescence

Heat Shock Alters Mesenchymal Stem Cell Identity and Induces Premature Senescence

The Analysis of In Vivo Aging in Human Bone Marrow Mesenchymal Stromal Cells Using Colony-Forming Unit-Fibroblast Assay and the CD45lowCD271+ Phenotype

Senescent mesenchymal stem/stromal cells and restoring their cellular functions

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Product

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The Analysis of In Vivo Aging in Human Bone Marrow Mesenchymal Stromal Cells Using Colony-Forming Unit-Fibroblast Assay and the CD45^lowCD271⁺ Phenotype