Modification of the alkaline comet assay with human mesenchymal stem cells

Cell Biology International

Volumn 36, Issue 1, 2012, Pages 113-117

  Fuchs, Robert ^a   Stelzer, Ingeborg ^a   Drees, Christoph M P ^a   Rehnolt, Christian ^a   Schraml, Elisabeth ^b   Sadjak, Anton ^a   Schwinger, Wolfgang ^a  

^a MEDICAL UNIVERSITY OF GRAZ   (Austria)

^b UNIVERSITY OF AGRICULTURAL SCIENCES   (Austria)

Author keywords

Comet assay; DNA damage; Human mesenchymal stem cell (MSC); replicative senescence

Indexed keywords

AGAROSE; DNA;

ARTICLE; CELL ADHESION; CELL DENSITY; CELL PROLIFERATION; COMET ASSAY; CONTROLLED STUDY; DNA DAMAGE; GROWTH RATE; HUMAN; HUMAN CELL; IMAGE ANALYSIS; IN VITRO STUDY; INTERMETHOD COMPARISON; MESENCHYMAL STEM CELL; OUTCOME ASSESSMENT; PROCESS OPTIMIZATION; REPRODUCIBILITY; SENESCENCE;

CELL AGING; CELL PROLIFERATION; CELLS, CULTURED; COMET ASSAY; DNA DAMAGE; HUMANS; MESENCHYMAL STEM CELLS; TELOMERE;

EID: 84858738051     PISSN: 10656995     EISSN: 10958355     Source Type: Journal    
DOI: 10.1042/CBI20110251     Document Type: Article

References (33)

1. Azqueta A, Herrmann K, Bading C, Meier S, Shaposhnikov S, Collins A. Towards a simpler, faster and higher capacity comet assay. In Proceedings of the Eighth International Comet Assay Workshop, Perugia, 2009.
2. Baptista J, Mercer C, Prigmore E, Gribble SM, Carter NP, Maloney V et al. Breakpoint mapping and array cgh in translocations: comparison of a phenotypically normal and an abnormal cohort. Am J Hum Genet 2008;82:927-36.
3. Bonab MM, Alimoghaddam K, Talebian F, Ghaffari SH, Ghavamzadeh A, Nikbin B. Aging of mesenchymal stem cell in vitro. BMC Cell Biol 2006;7:14.
4. Burhans WC, Weinberger M. DNA replication stress, genome instability and aging. Nucleic Acids Res 2007;35:7545-56.
5. Cao X, Liu M, Tuo J, Shen D, Chan CC. The effects of quercetin in cultured human rpe cells under oxidative stress and in ccl2/cx3cr1 double deficient mice. Exp Eye Res 2010;9:15-25.
6. Chaubey RC. Computerized image analysis software for the comet assay. Methods Mol Biol 2005;29:97-106.
7. Collins AR. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 2004;26:249-61.
8. Dusinska M, Collins AR. The comet assay in human biomonitoring: gene- environment interactions. Mutagenesis 2008;23:191-205.
9. Fehrer C, Lepperdinger G. Mesenchymal stem cell aging. Exp Gerontol 2005;40:926-30.
10. Galderisi U, Helmbold H, Squillaro T, Alessio N, Komm N, Khadang B et al. In vitro senescence of rat mesenchymal stem cells is accompanied by downregulation of stemness-related and DNA damage repair genes. Stem Cells Dev 2009;18:1033-42.
11. Hinkal GW, Gatza CE, Parikh N, Donehower LA. Altered senescence, apoptosis, and DNA damage response in a mutant p53 model of accelerated aging. Mech Ageing Dev 2009;130:262-71.
12. Klaude M, Eriksson S, Nygren J, Ahnstrom G. The comet assay: mechanisms and technical considerations. Mutat Res 1996;363:89-96.
13. Mughal A, Vikram A, Ramarao P, Jena GB. Micronucleus and comet assay in the peripheral blood of juvenile rat: establishment of assay feasibility, time of sampling and the induction of DNA damage. Mutat Res 2010;700:86-94.
14. Nousis L, Doulias PT, Aligiannis N, Bazios D, Agalias A, Galaris D et al. DNA protecting and genotoxic effects of olive oil related components in cells exposed to hydrogen peroxide. Free Radical Res 2005;39:787-95.
15. Olive PL. The comet assay. An overview of techniques. In: Didenko VV, editor. Series: Methods in Molecular Biology, vol. 203: In Situ Detection of DNA Damage: Methods and Protocols. Totowa: Humana Press; 2002. p. 179-94.
16. Olive PL, Banath JP. The comet assay: a method to measure DNA damage in individual cells. Nat Protoc 2006;1:23-9.
17. Ostling O, Johanson KJ. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem Biophys Res Commun 1984;123:291-8.
18. Passos JF, Von Zglinicki T. Oxygen free radicals in cell senescence: are they signal transducers? Free Radical Res 2006;40:1277-83.
19. Perez-Simon JA, Tabera S, Sarasquete ME, Diez-Campelo M, Canchado J, Sanchez-Abarca LI et al. Mesenchymal stem cells are functionally abnormal in patients with immune thrombocytopenic purpura. Cytotherapy 2009;11:698-705.
20. Pfragner R, Behmel A, Hoger H, Beham A, Ingolic E, Stelzer I et al. Establishment and characterization of three novel cell lines-p-sts, l-sts, h-sts-derived from a human metastatic midgut carcinoid. Anticancer Res 2009;29:1951-61.
21. Reinisch A, Strunk D. Isolation and animal serum free expansion of human umbilical cord derived mesenchymal stromal cells (MSCs) and endothelial colony forming progenitor cells (ECFCs). J Vis Exp 2009;32:1525.
22. Sadan O, Melamed E, Offen D. Bone-marrow-derived mesenchymal stem cell therapy for neurodegenerative diseases. Expert Opin Biol Ther 2009;9:1487-97.
23. Schallmoser K, Rohde E, Reinisch A, Bartmann C, Thaler D, Drexler C et al. Rapid large-scale expansion of functional mesenchymal stem cells from unmanipulated bone marrow without animal serum. Tissue Eng Part C Methods 2008;14:185-96.
24. Schallmoser K, Bartmann C, Rohde E, Bork S, Guelly C, Obenauf AC et al. Replicative senescence-associated gene expression changes in mesenchymal stromal cells are similar under different culture conditions. Haematologica 2010;95:867-74.
25. Schraml E, Quan P, Stelzer I, Fuchs R, Skalicky M, Viidik A et al. Norepinephrine treatment and aging lead to systemic and intracellular oxidative stress in rats. Exp Gerontol 2007;42:1072-8.
26. Schraml E, Fuchs R, Kotzbeck P, Grillari J, Schauenstein K. Acute adrenergic stress inhibits proliferation of murine haematopoietic progenitor cells via p38/mapk signaling. Stem Cells Dev 2009;18:215-27.
27. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988;175:184-91.
28. Thomas S, Green MH, Lowe JE, Green IC. Measurement of DNA damage using the comet assay. In: Titheradge MA, editor. Series: Methods in Molecular Biology, vol. 100: Nitric Oxide Protocols. Totowa: Humana Press; 1998. p. 301-10.
29. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 2000;35:206-21.
30. Wagner W, Horn P, Castoldi M, Diehlmann A, Bork S, Saffrich R et al. Replicative senescence of mesenchymal stem cells: a continuous and organized process. PLoS ONE 2008;3:e2213.
31. Wagner W, Bork S, Lepperdinger G, Joussen S, Ma N, Strunk D et al. How to track cellular aging of mesenchymal stromal cells? Aging (Albany NY) 2010;2:224-30.
32. Whone AL, Scolding NJ. Mesenchymal stem cells and neurodegenerative disease. Clin Pharmacol Ther 2009;85:19-20.
33. Zheng J, Edelman SW, Tharmarajah G, Walker DW, Pletcher SD, Seroude L. Differential patterns of apoptosis in response to aging in drosophila. Proc Natl Acad Sci U.S.A 2005;102:12083-88.