Angiogenic properties of aged adipose derived mesenchymal stem cells after hypoxic conditioning.
BACKGROUND
Mesenchymal stem cells derived from adipose tissue (ADSC) are multipotent stem cells, originated from the vascular-stromal compartment of fat tissue. ADSC are used as an alternative cell source for many different cell therapies, however in ischemic cardiovascular diseases the therapeutic benefit was modest. One of the reasons could be the use of autologous aged ADSC, which recently were found to have impaired functions. We therefore analysed the effects of age on age markers and angiogenic properties of ADSC. Hypoxic conditioning was investigated as a form of angiogenic stimulation.


METHODS
ADSC were harvested from young (1-3 month), adult (12 month) and aged (18-24 month) mice and cultured under normoxic (20%) and hypoxic (1%) conditions for 48 h. Differences in proliferation, apoptosis and telomere length were assessed in addition to angiogenic properties of ADSC.


RESULTS
Proliferation potential and telomere length were decreased in aged ADSC compared to young ADSC. Frequency of apoptotic cells was higher in aged ADSC. Gene expression of pro-angiogenic factors including vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and hepatic growth factor (HGF) were down-regulated with age, which could be restored by hypoxia. Transforming growth factor (TGF-β) increased in the old ADSC but was reduced by hypoxia.Expression of anti-angiogenic factors including thrombospondin-1 (TBS1) and plasminogen activator inhibitor-1 (PAI-1) did increase in old ADSC, but could be reduced by hypoxic stimulation. Endostatin (ENDS) was the highest in aged ADSC and was also down-regulated by hypoxia. We noted higher gene expression of proteases system factors like urokinase-type plasminogen activator receptor (uPAR), matrix metalloproteinases (MMP2 and MMP9) and PAI-1 in aged ADSC compared to young ADSC, but they decreased in old ADSC. Tube formation on matrigel was higher in the presence of conditioned medium from young ADSC in comparison to aged ADSC.


CONCLUSIONS
ADSC isolated from older animals show changes, including impaired proliferation and angiogenic stimulation. Angiogenic gene expression can be partially be improved by hypoxic preconditioning, however the effect is age-dependent. This supports the hypothesis that autologous ADSC from aged subjects might have an impaired therapeutic potential.
Department of Biological and Medical Chemistry, Faculty of Fundamental Medicine, Lomonosov Moscow State University, Moscow, Russia.
Anastasia
A
Efimenko
Ekaterina
E
Starostina
Natalia
N
Kalinina
Alexandra
A
Stolzing
D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D000375:Aging / Q000502:physiology; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D015687:Cell Hypoxia / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism
D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D010100:Oxygen
Stem Cells. 2007 May;25(5):1166-77
17289933
J Biomed Sci. 2006 Jan;13(1):47-58
16283432
Ageing Res Rev. 2006 Feb;5(1):91-116
16310414
Curr Opin Lipidol. 2007 Oct;18(5):508-14
17885420
Stem Cells Dev. 2006 Aug;15(4):478-87
16978051
FEBS Lett. 1998 Dec 4;440(3):399-402
9872410
Aging Cell. 2007 Dec;6(6):745-57
17925003
Am J Geriatr Cardiol. 2001 Nov-Dec;10(6):348-54
11684920
Cell. 1997 Jan 24;88(2):277-85
9008168
Arterioscler Thromb Vasc Biol. 2009 Jan;29(1):61-6
18974384
Tissue Eng Part A. 2009 Aug;15(8):2039-50
19368510
Surgery. 2009 Aug;146(2):190-7
19628073
Am J Physiol Regul Integr Comp Physiol. 2009 Nov;297(5):R1563-9
19776253
Immunol Lett. 2003 Oct 31;89(2-3):267-70
14556988
Trends Biotechnol. 2006 Apr;24(4):150-4
16488036
Keio J Med. 2005 Sep;54(3):132-41
16237275
Cancer Res. 2005 Apr 15;65(8):3035-9
15833829
Tissue Eng. 2001 Apr;7(2):211-28
11304456
Circulation. 2003 Jan 21;107(2):230-3
12538420
Nucleic Acids Res. 2002 May 15;30(10):e47
12000852
J Cardiovasc Med (Hagerstown). 2010 Feb;11(2):71-80
19996982
BMC Cell Biol. 2007 May 29;8:18
17535427
Stem Cells. 2008 Oct;26(10):2625-33
18669910
J Cell Physiol. 2008 Feb;214(2):413-21
17654479
Brain Res Dev Brain Res. 2003 Oct 10;145(1):141-9
14519500
Biochem Biophys Res Commun. 1998 May 8;246(1):255-60
9600102
Mol Biol Cell. 2005 Mar;16(3):1131-41
15635089
Aging Cell. 2009 Dec;8(6):676-89
19732043
Free Radic Biol Med. 1997;23(3):361-6
9214571
Pharmacology. 1999 Apr;58(4):200-8
10077739
Circ Res. 2007 May 11;100(9):1249-60
17495232
Mech Ageing Dev. 2008 Mar;129(3):163-73
18241911
Sci Aging Knowledge Environ. 2004 Feb 18;2004(7):pe7
14973174
J Am Coll Cardiol. 2008 Apr 8;51(14):1384-92
18387441
Cytokine. 2009 Apr;46(1):100-3
19223198
Mech Ageing Dev. 1996 Dec 20;92(2-3):121-32
9080393
Am J Physiol Cell Physiol. 2008 Nov;295(5):C1271-80
18787077
Stem Cells. 2007 Apr;25(4):818-27
17420225
Best Pract Res Clin Haematol. 2004 Sep;17(3):387-99
15498711
Proc Inst Med Chic. 1975 Jul-Sep;30(8):256-8
1103123
Stem Cells. 2004;22(5):675-82
15342932
Bone. 2007 Apr;40(4):1078-87
17276151
Connect Tissue Res. 1996;35(1-4):93-9
9084647
Biomed Mater Eng. 2008;18(1 Suppl):S71-6
18334717
J Mol Cell Cardiol. 2008 Mar;44(3):582-96
18199451
Fertil Steril. 2004 Sep;82(3):756-9
15374730
J Dermatol Sci. 2007 Oct;48(1):43-52
17644316
J Biol Chem. 2000 Sep 22;275(38):29643-7
10882714
J Histochem Cytochem. 2003 Sep;51(9):1119-30
12923237
Cytotherapy. 2006;8(4):315-7
16923606
Circulation. 2004 Mar 16;109(10):1292-8
14993122
Mol Biol Cell. 2002 Dec;13(12):4279-95
12475952
Stem Cells. 2003;21(3):337-47
12743328
Nat Cell Biol. 2006 Aug;8(8):877-84
16862142
Cells Tissues Organs. 2009;190(2):81-93
19033681
Lancet. 2003 Jan 4;361(9351):45-6
12517467
Arterioscler Thromb Vasc Biol. 2005 Dec;25(12):2542-7
16224047
J Cell Mol Med. 2008 Apr;12(2):553-63
18419597
J Biol Chem. 2007 Nov 30;282(48):34938-44
17913706
Circ Res. 2008 Jan 4;102(1):77-85
17967785
Curr Pharm Des. 2009;15(24):2784-90
19689349
Aging Cell. 2008 Jun;7(3):335-43
18248663
Nat Biotechnol. 2004 May;22(5):560-7
15077117
Tsitologiia. 2010;52(2):144-54
20352697
J Thorac Cardiovasc Surg. 2008 Apr;135(4):799-808
18374759
Biochem Biophys Res Commun. 2003 Feb 21;301(4):1016-22
12589814
Curr Drug Targets. 2005 Feb;6(1):31-41
15720211
J Cell Biochem. 2006 Aug 1;98(5):1076-84
16619257