The regenerative potential diminishes with age and this has been ascribed to functional impairments of adult stem cells. Cells in culture undergo senescence after a certain number of cell divisions whereby the cells enlarge and finally stop proliferation. This observation of replicative senescence has been extrapolated to somatic stem cells in vivo and might reflect the aging process of the whole organism. In this study we have analyzed the effect of aging on gene expression profiles of human mesenchymal stromal cells (MSC) and human hematopoietic progenitor cells (HPC). MSC were isolated from bone marrow of donors between 21 and 92 years old. 67 genes were age-induced and 60 were age-repressed. HPC were isolated from cord blood or from mobilized peripheral blood of donors between 27 and 73 years and 432 genes were age-induced and 495 were age-repressed. The overlap of age-associated differential gene expression in HPC and MSC was moderate. However, it was striking that several age-related gene expression changes in both MSC and HPC were also differentially expressed upon replicative senescence of MSC in vitro. Especially genes involved in genomic integrity and regulation of transcription were age-repressed. Although telomerase activity and telomere length varied in HPC particularly from older donors, an age-dependent decline was not significant arguing against telomere exhaustion as being causal for the aging phenotype. These studies have demonstrated that aging causes gene expression changes in human MSC and HPC that vary between the two different cell types. Changes upon aging of MSC and HPC are related to those of replicative senescence of MSC in vitro and this indicates that our stem and progenitor cells undergo a similar process also in vivo.
References
[1]
Ho AD, Wagner W, Mahlknecht U (2005) Stem cells and ageing. The potential of stem cells to overcome age-related deteriorations of the body in regenerative medicine. EMBO Rep 6: 35–38.
[2]
Gazit R, Weissman IL, Rossi DJ (2008) Hematopoietic stem cells and the aging hematopoietic system. Semin Hematol 45: 218–224.
[3]
Giangreco A, Qin M, Pintar JE, Watt FM (2008) Epidermal stem cells are retained in vivo throughout skin aging. Aging Cell 7: 250–259.
[4]
Horwitz EM, Le BK, Dominici M, Mueller I, Slaper-Cortenbach I, et al. (2005) Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement. Cytotherapy 7: 393–395.
[5]
Horn P, Bork S, Diehlmann A, Walenda T, Eckstein V, et al. (2008) Isolation of human mesenchymal stromal cells is more efficient by red blood cell lysis. Cytotherapy 10: 676–685.
[6]
Ho AD, Wagner W, Franke W (2008) Heterogeneity of mesenchymal stromal cell preparations. Cytotherapy 10: 320–330.
[7]
Wagner W, Ho AD (2007) Mesenchymal stem cell preparations-comparing apples and oranges. Stem Cell Rev 3: 239–248.
[8]
von Zglinicki T, Martin-Ruiz CM (2005) Telomeres as biomarkers for ageing and age-related diseases. Curr Mol Med 5: 197–203.
[9]
Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 25: 585–621.
[10]
Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37: 614–636.
[11]
Smith JR, Pereira-Smith OM (1996) Replicative senescence: implications for in vivo aging and tumor suppression. Science 273: 63–67.
[12]
Bonab MM, Alimoghaddam K, Talebian F, Ghaffari SH, Ghavamzadeh A, et al. (2006) Aging of mesenchymal stem cell in vitro. BMC Cell Biol 7: 14.
[13]
Stenderup K, Justesen J, Clausen C, Kassem M (2003) Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone 33: 919–926.
Baxter MA, Wynn RF, Jowitt SN, Wraith JE, Fairbairn LJ, et al. (2004) Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion. Stem Cells 22: 675–682.
[16]
Wagner W, Horn P, Castoldi M, Diehlmann A, Bork S, et al. (2008) Replicative Senescence of Mesenchymal Stem Cells - a Continuous and Organized Process. PLoS ONE 5: e2213.
[17]
Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, et al. (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8: 315–317.
[18]
Kern S, Eichler H, Stoeve J, Kluter H, Bieback K (2006) Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells 24: 1294–1301.
[19]
Fehrer C, Brunauer R, Laschober G, Unterluggauer H, Reitinger S, et al. (2007) Reduced oxygen tension attenuates differentiation capacity of human mesenchymal stem cells and prolongs their lifespan. Aging Cell 6: 745–757.
[20]
Bieback K, Kern S, Kluter H, Eichler H (2004) Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood. Stem Cells 22: 625–634.
[21]
Csoka AB, English SB, Simkevich CP, Ginzinger DG, Butte AJ, et al. (2004) Genome-scale expression profiling of Hutchinson-Gilford progeria syndrome reveals widespread transcriptional misregulation leading to mesodermal/mesenchymal defects and accelerated atherosclerosis. Aging Cell 3: 235–243.
[22]
Cobb J, Dierich A, Huss-Garcia Y, Duboule D (2006) A mouse model for human short-stature syndromes identifies Shox2 as an upstream regulator of Runx2 during long-bone development. Proc Natl Acad Sci U S A 103: 4511–4515.
[23]
Scaffidi P, Misteli T (2006) Lamin A-dependent nuclear defects in human aging. Science 312: 1059–1063.
[24]
Assounga AG, Warner CM (2004) Transcription of major histocompatibility complex class I (Kb) and transporter associated with antigen processing 1 and 2 genes is up-regulated with age. Immunology 113: 378–383.
[25]
Janick-Buckner D, Briggs CJ, Meyer TE, Harvey N, Warner CM (1991) Major histocompatibility complex antigen expression on lymphocytes from aging strain A mice. Growth Dev Aging 55: 53–62.
[26]
Walenda T, Bork S, Horn P, Wein F, Saffrich R, et al. (2009) Co-Culture with Mesenchymal Stromal Cells Increases Proliferation and Maintenance of Hematopoietic Progenitor Cells. J Cell Mol Med. in print.
[27]
Henry RW, Mittal V, Ma B, Kobayashi R, Hernandez N (1998) SNAP19 mediates the assembly of a functional core promoter complex (SNAPc) shared by RNA polymerases II and III. Genes Dev 12: 2664–2672.
[28]
Weng N (2001) Interplay between telomere length and telomerase in human leukocyte differentiation and aging. J Leukoc Biol 70: 861–867.
[29]
Nalapareddy K, Jiang H, Guachalla Gutierrez LM, Rudolph KL (2008) Determining the influence of telomere dysfunction and DNA damage on stem and progenitor cell aging: what markers can we use? Exp Gerontol 43: 998–1004.
[30]
Krunic D, Moshir S, Greulich-Bode KM, Figueroa R, Cerezo A, et al. (2009) Tissue context-activated telomerase in human epidermis correlates with littleage-dependent telomere loss. Biochimica et Biophysica Acta 1792: 297–208.
[31]
Cerezo A, Stark HJ, Moshir S, Boukamp P (2003) Constitutive overexpression of human telomerase reverse transcriptase but not c-myc blocks terminal differentiation in human HaCaT skin keratinocytes. J Invest Dermatol 121: 110–119.
[32]
de Haan G (2007) Epigenetic control of hematopoietic stem cell aging, the case of Ezh2. Ann N Y Acad Sci Epub. ahead of print.
[33]
Chambers SM, Shaw CA, Gatza C, Fisk CJ, Donehower LA, et al. (2007) Aging hematopoietic stem cells decline in function and exhibit epigenetic dysregulation. PLoS Biol 5: 201.
[34]
Rossi DJ, Bryder D, Zahn JM, Ahlenius H, Sonu R, et al. (2005) Cell intrinsic alterations underlie hematopoietic stem cell aging. Proc Natl Acad Sci U S A 102: 9194–9199.
[35]
Prall WC, Czibere A, Jager M, Spentzos D, Libermann TA, et al. (2007) Age-related transcription levels of KU70, MGST1 and BIK in CD34+ hematopoietic stem and progenitor cells. Mech Ageing Dev 128: 503–510.
[36]
Hayflick L (2007) Biological aging is no longer an unsolved problem. Ann N Y Acad Sci 1100: 1–13.
[37]
Kirkwood TB (1977) Evolution of ageing. Nature 270: 301–304.
[38]
Kirkwood TB (2008) Understanding ageing from an evolutionary perspective. J Intern Med 263: 117–127.
[39]
Janzen V, Forkert R, Fleming HE, Saito Y, Waring MT, et al. (2006) Stem-cell ageing modified by the cyclin-dependent kinase inhibitor p16INK4a. Nature 443: 421–426.
[40]
Kiyono T, Foster SA, Koop JI, McDougall JK, Galloway DA, et al. (1998) Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature 396: 84–88.
[41]
Dumble M, Moore L, Chambers SM, Geiger H, Van Zant G, et al. (2007) The impact of altered p53 dosage on hematopoietic stem cell dynamics during aging. Blood 109: 1736–1742.
[42]
Kamminga LM, de Haan G (2006) Cellular memory and hematopoietic stem cell aging. Stem Cells 24: 1143–1149.
[43]
Noer A, Boquest AC, Collas P (2007) Dynamics of adipogenic promoter DNA methylation during clonal culture of human adipose stem cells to senescence. BMC Cell Biol 8: 18.
[44]
Schneider EL, Mitsui Y (1976) The relationship between in vitro cellular aging and in vivo human age. Proc Natl Acad Sci U S A 73: 3584–3588.
[45]
Mareschi K, Ferrero I, Rustichelli D, Aschero S, Gammaitoni L, et al. (2006) Expansion of mesenchymal stem cells isolated from pediatric and adult donor bone marrow. J Cell Biochem 97: 744–754.
[46]
Cristofalo VJ, Allen RG, Pignolo RJ, Martin BG, Beck JC (1998) Relationship between donor age and the replicative lifespan of human cells in culture: a reevaluation. Proc Natl Acad Sci U S A 95: 10614–10619.
[47]
Goldstein S, Moerman EJ, Soeldner JS, Gleason RE, Barnett DM (1978) Chronologic and physiologic age affect replicative life-span of fibroblasts from diabetic, prediabetic, and normal donors. Science 199: 781–782.
[48]
Colter DC, Class R, DiGirolamo CM, Prockop DJ (2000) Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci U S A 97: 3213–3218.
[49]
Breault DT, Min IM, Carlone DL, Farilla LG, Ambruzs DM, et al. (2008) Generation of mTert-GFP mice as a model to identify and study tissue progenitor cells. Proc Natl Acad Sci U S A 105: 10420–10425.
[50]
Morrison SJ, Prowse KR, Ho P, Weissman IL (1996) Telomerase activity in hematopoietic cells is associated with self-renewal potential. Immunity 5: 207–216.
[51]
Vaziri H, Dragowska W, Allsopp RC, Thomas TE, Harley CB, et al. (1994) Evidence for a mitotic clock in human hematopoietic stem cells: loss of telomeric DNA with age. Proc Natl Acad Sci U S A 91: 9857–9860.
[52]
O'Hare MJ, Bond J, Clarke C, Takeuchi Y, Atherton AJ, et al. (2001) Conditional immortalization of freshly isolated human mammary fibroblasts and endothelial cells. Proc Natl Acad Sci U S A 98: 646–651.
[53]
Di Donna S, Mamchaoui K, Cooper RN, Seigneurin-Venin S, Tremblay J, et al. (2003) Telomerase can extend the proliferative capacity of human myoblasts, but does not lead to their immortalization. Mol Cancer Res 1: 643–653.
[54]
Zimmermann S, Glaser S, Ketteler R, Waller CF, Klingmuller U, et al. (2004) Effects of telomerase modulation in human hematopoietic progenitor cells. Stem Cells 22: 741–749.
[55]
Ju Z, Jiang H, Jaworski M, Rathinam C, Gompf A, et al. (2007) Telomere dysfunction induces environmental alterations limiting hematopoietic stem cell function and engraftment. Nat Med 13: 742–747.
[56]
Wagner W, Ansorge A, Wirkner U, Eckstein V, Schwager C, et al. (2004) Molecular evidence for stem cell function of the slow-dividing fraction among human hematopoietic progenitor cells by genome-wide analysis. Blood 104: 675–686.
[57]
Ito K, Hirao A, Arai F, Takubo K, Matsuoka S, et al. (2006) Reactive oxygen species act through p38 MAPK to limit the lifespan of hematopoietic stem cells. Nat Med 12: 446–451.
[58]
Ogden DA, Mickliem HS (1976) The fate of serially transplanted bone marrow cell populations from young and old donors. Transplantation 22: 287–293.
[59]
Allsopp RC, Morin GB, Horner JW, DePinho R, Harley CB, et al. (2003) Effect of TERT over-expression on the long-term transplantation capacity of hematopoietic stem cells. Nat Med 9: 369–371.
[60]
Allsopp RC, Cheshier S, Weissman IL (2001) Telomere shortening accompanies increased cell cycle activity during serial transplantation of hematopoietic stem cells. J Exp Med 193: 917–924.
[61]
Wilson A, Laurenti E, Oser G, van der Wath RC, Blanco-Bose W, et al. (2008) Hematopoietic stem cells reversibly switch from dormancy to self-renewal during homeostasis and repair. Cell 135: 1118–1129.
[62]
Wagner W, Horn P, Bork S, Ho AD (2008) Aging of hematopoietic stem cells is regulated by the stem cell niche. Experimental Gerontology 43: 974–980.
[63]
Kim M, Moon HB, Spangrude GJ (2003) Major age-related changes of mouse hematopoietic stem/progenitor cells. Ann N Y Acad Sci 996: 195–208.
[64]
Dykstra B, de Haan G (2008) Hematopoietic stem cell aging and self-renewal. Cell Tissue Res 331: 91–101.
[65]
Lee HS, Huang GT, Chiang H, Chiou LL, Chen MH, et al. (2003) Multipotential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis. Stem Cells 21: 190–199.
[66]
Wagner W, Wein F, Seckinger A, Frankhauser M, Wirkner U, et al. (2005) Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp Hematol 33: 1402–1416.
[67]
Wagner W, Feldmann RE Jr, Seckinger A, Maurer MH, Wein F, et al. (2006) The heterogeneity of human mesenchymal stem cell preparations-Evidence from simultaneous analysis of proteomes and transcriptomes. Exp Hematol 34: 536–548.
[68]
Reyes M, Lund T, Lenvik T, Aguiar D, Koodie L, et al. (2001) Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98: 2615–2625.
[69]
Haynesworth SE, Baber MA, Caplan AI (1992) Cell surface antigens on human marrow-derived mesenchymal cells are detected by monoclonal antibodies. Bone 13: 69–80.
[70]
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, et al. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–147.
[71]
Bolstad BM, Irizarry RA, Astrand M, Speed TP (2003) A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19: 185–193.
[72]
Saeed AI, Sharov V, White J, Li J, Liang W, et al. (2003) TM4: a free, open-source system for microarray data management and analysis. Biotechniques 34: 374–378.
[73]
Pavlidis P, Noble WS (2001) Analysis of strain and regional variation in gene expression in mouse brain. Genome Biol 2: 42.
[74]
Blake J, Schwager C, Kapushesky M, Brazma A (2006) ChroCoLoc: an application for calculating the probability of co-localization of microarray gene expression. Bioinformatics 22: 765–767.
[75]
Harle-Bachor C, Boukamp P (1996) Telomerase activity in the regenerative basal layer of the epidermis inhuman skin and in immortal and carcinoma-derived skin keratinocytes. Proc Natl Acad Sci U S A 93: 6476–6481.
