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Characterization of senescent mesenchymal stem/stromal cells derived from equine bone marrow and the effects of NANOG on the senescent phenotypes.
Abstract: In equine regenerative medicine using bone marrow-derived mesenchymal stem/stromal cells (BM-MSC), the importance of the quality management of BM-MSC has been widely recognized. However, there is little information concerning the relationship between cellular senescence and the stemness in equine BM-MSC. In this study, we showed that stemness markers (NANOG, OCT4, SOX2 and telomerase reverse transcriptase) and colony forming unit-fibroblast apparently decreased accompanied with incidence of senescence-associated β-galactosidase-positive cells by repeated passage. Additionally, we suggested that down-regulation of cell proliferation in senescent BM-MSC was related to increased expression of cyclin-dependent kinase inhibitor 2B (CDKN2B). On the other hand, forced expression of NANOG into senescent BM-MSC brought upregulation of several stemness markers and downregulation of CKDN2B accompanied with restoration of proliferation potential and osteogenic ability. These results suggested that expression of NANOG was important for the maintenance of the stemness in equine BM-MSC.
Publication Date: 2024-07-08 PubMed ID: 38972751PubMed Central: PMC11422694DOI: 10.1292/jvms.24-0161Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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Overview
- This study investigated how aging (senescence) affects mesenchymal stem/stromal cells (BM-MSC) derived from horse bone marrow, focusing on the relationship between cellular aging and stem cell characteristics (“stemness”).
- The researchers also evaluated the role of the gene NANOG in reversing senescent traits and preserving the regenerative capacity of these cells.
Background and Purpose
- Equine regenerative medicine employs BM-MSCs to repair tissues; the therapeutic efficacy depends on the quality and functionality of these stem cells.
- Cellular senescence, or the process by which cells cease to divide and lose function over time, is a barrier to effective stem cell therapies but its relationship to stemness in horse BM-MSC is not well understood.
- The study aimed to characterize senescent BM-MSC and understand how stemness markers change with cell aging.
- It further sought to determine whether overexpression of NANOG, a key stemness gene, could rejuvenate aged BM-MSCs.
Key Methods
- Equine BM-MSCs were cultured through multiple passages to induce senescence.
- Markers of stemness (NANOG, OCT4, SOX2, telomerase reverse transcriptase) and senescence (β-galactosidase positivity) were measured at different cell passages.
- Proliferation was assessed by colony-forming unit-fibroblast (CFU-F) assays.
- Expression of cell cycle inhibitors like cyclin-dependent kinase inhibitor 2B (CDKN2B) was evaluated to understand proliferation decline.
- NANOG was forcibly expressed in senescent BM-MSCs using genetic techniques to assess its effects on stemness markers, cell proliferation, and ability to differentiate into bone cells (osteogenic potential).
Results
- Repeated cell culture passages led to increased cellular senescence, demonstrated by more β-galactosidase positive cells.
- With senescence, stemness markers including NANOG, OCT4, SOX2, and telomerase reverse transcriptase significantly decreased, correlating with reduced clonogenic capacity (CFU-F).
- The decline in cell proliferation was associated with upregulation of CDKN2B, a protein known to arrest the cell cycle.
- Introducing NANOG into senescent BM-MSCs reversed these senescent features by:
- Increasing expression of multiple stemness markers.
- Decreasing CDKN2B levels, thus lifting cell cycle inhibition.
- Restoring the proliferative ability of the cells.
- Regaining the cells’ capacity to differentiate into bone-forming cells, indicating functional rejuvenation.
Conclusions and Implications
- Senescence in equine BM-MSCs leads to reduced stemness and proliferation, limiting their therapeutic potential.
- NANOG plays a critical role in maintaining stemness and mitigating senescence-related decline in these cells.
- Forced expression of NANOG could be a promising strategy to rejuvenate aged or extensively cultured BM-MSCs for improved outcomes in equine regenerative medicine.
- This work enhances understanding of molecular changes during BM-MSC aging and highlights potential genetic interventions to preserve stem cell function.
Cite This Article
APA
Kushida C, Tamura N, Kasashima Y, Sato K, Arai K.
(2024).
Characterization of senescent mesenchymal stem/stromal cells derived from equine bone marrow and the effects of NANOG on the senescent phenotypes.
J Vet Med Sci, 86(9), 930-937.
https://doi.org/10.1292/jvms.24-0161 Publication
Researcher Affiliations
- Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo, Japan.
- National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, Tokyo, Japan.
- Laboratory of Clinical Science and Pathobiology, Equine Research Institute, Japan Racing Association, Tochigi, Japan.
- Laboratory of Clinical Science and Pathobiology, Equine Research Institute, Japan Racing Association, Tochigi, Japan.
- National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, Tokyo, Japan.
- Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo, Japan.
MeSH Terms
- Animals
- Mesenchymal Stem Cells / physiology
- Mesenchymal Stem Cells / cytology
- Horses
- Cellular Senescence / physiology
- Nanog Homeobox Protein / metabolism
- Nanog Homeobox Protein / genetics
- Bone Marrow Cells / physiology
- Bone Marrow Cells / cytology
- Cell Proliferation
- Phenotype
Conflict of Interest Statement
The authors declare that they have no competing interests.
References
This article includes 21 references
- Batsali AK, Pontikoglou C, Koutroulakis D, Pavlaki KI, Damianaki A, Mavroudi I, Alpantaki K, Kouvidi E, Kontakis G, Papadaki HA. Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton’s jelly and bone marrow-derived mesenchymal stem cells.. 2017;8:102.
- Eto S, Goto M, Soga M, Kaneko Y, Uehara Y, Mizuta H, Era T. Mesenchymal stem cells derived from human iPS cells via mesoderm and neuroepithelium have different features and therapeutic potentials.. 2018;13:e0200790.
- Flores I, Benetti R, Blasco MA. Telomerase regulation and stem cell behaviour.. 2006;18:254–260.
- Go MJ, Takenaka C, Ohgushi H. Forced expression of Sox2 or Nanog in human bone marrow derived mesenchymal stem cells maintains their expansion and differentiation capabilities.. 2008;314:1147–1154.
- Godwin EE, Young NJ, Dudhia J, Beamish IC, Smith RKW. Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon.. 2012;44:25–32.
- Itahana K, Itahana Y, Dimri GP. Colorimetric detection of senescence-associated β galactosidase.. 2013;965:143–156.
- Kasashima Y, Ueno T, Tomita A, Goodship AE, Smith RK. Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells.. 2011;43:288–294.
- Liu L, DiGirolamo CM, Navarro PAAS, Blasco MA, Keefe DL. Telomerase deficiency impairs differentiation of mesenchymal stem cells.. 2004;294:1–8.
- Liu J, Ding Y, Liu Z, Liang X. Senescence in mesenchymal stem cells: Functional slterations, molecular mechanisms, and rejuvenation strategies.. 2020;8:258.
- Lu H, Lyu Y, Tran L, Lan J, Xie Y, Yang Y, Murugan NL, Wang YJ, Semenza GL. HIF-1 recruits NANOG as a coactivator for TERT gene transcription in hypoxic breast cancer stem cells.. 2021;36:109757.
- O’Connor K. A cautionary tale about the use of colony-forming efficiency as a proxy for the survival of mesenchymal stem cells.. 2020;11:292.
- Roche S, Richard MJ, Favrot MC. Oct-4, Rex-1, and Gata-4 expression in human MSC increase the differentiation efficiency but not hTERT expression.. 2007;101:271–280.
- Sasao T, Fukuda Y, Yoshida S, Miyabara S, Kasashima Y, Kuwano A, Arai K. Population doubling level-dependent change of secreted glycosaminoglycan in equine bone marrow-derived mesenchymal stem cells.. 2015;26:73–80.
- Shahini A, Mistriotis P, Asmani M, Zhao R, Andreadis ST. Nanog restores contractility of mesenchymal stem cell-based senescent microtissues.. 2017;23:535–545.
- Simonsen JL, Rosada C, Serakinci N, Justesen J, Stenderup K, Rattan SIS, Jensen TG, Kassem M. Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells.. 2002;20:592–596.
- Squillaro T, Severino V, Alessio N, Farina A, Di Bernardo G, Cipollaro M, Peluso G, Chambery A, Galderisi U. De-regulated expression of the BRG1 chromatin remodeling factor in bone marrow mesenchymal stromal cells induces senescence associated with the silencing of NANOG and changes in the levels of chromatin proteins.. 2015;14:1315–1326.
- Tsai CC, Chen CL, Liu HC, Lee YT, Wang HW, Hou LT, Hung SC. Overexpression of hTERT increases stem-like properties and decreases spontaneous differentiation in human mesenchymal stem cell lines.. 2010;17:64.
- Tsai CC, Su PF, Huang YF, Yew TL, Hung SC. Oct4 and Nanog directly regulate Dnmt1 to maintain self-renewal and undifferentiated state in mesenchymal stem cells.. 2012;47:169–182.
- Vidal MA, Walker NJ, Napoli E, Borjesson DL. Evaluation of senescence in mesenchymal stem cells isolated from equine bone marrow, adipose tissue, and umbilical cord tissue.. 2012;21:273–283.
- Wexler SA, Donaldson C, Denning-Kendall P, Rice C, Bradley B, Hows JM. Adult bone marrow is a rich source of human mesenchymal ‘stem’ cells but umbilical cord and mobilized adult blood are not.. 2003;121:368–374.
- Zahedi M, Parham A, Dehghani H, Kazemi Mehrjerdi H. Equine bone marrow-derived mesenchymal stem cells: optimization of cell density in primary culture.. 2018;5:31.
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