Expansion of adipose tissue‐derived stromal cells at “physiologic” hypoxia attenuates replicative senescence

Abstract

Multipotent mesenchymal stromal cells are considered as a perspective tool in cell therapy and regenerative medicine. Unfortunately, autologous cell therapy does not always provide positive outcomes in elder donors, perhaps as a result of the alterations of stem cell compartments. The mechanisms of stem and progenitor cell senescence and the factors engaged are investigated intensively. In present paper, we elucidated the effects of tissue-related O on morphology, functions, and transcriptomic profile of adipose tissue-derived stromal cells (ASCs) in replicative senescence in vitro model. Replicatively senescent ASCs at ambient (20%) O (12-21 passages) demonstrated an increased average cell size, granularity, reactive oxygen species level, including anion superoxide, lysosomal compartment activity, and IL-6 production. Decreased ASC viability and proliferation, as well as the change of more than 10 senescence-associated gene expression were detected (IGF1, CDKN1C, ID1, CCND1, etc). Long-term ASC expansion at low O (5%) revoked in part the replicative senescence-associated alterations.

Identification of new genes associated to senescent and tumorigenic phenotypes in mesenchymal stem cells

“…Among the 18 upregulated, 6 genes ( ANKRD1 , NTN4 , OXTR , SERPINB2 , SYNPO2 and THBS1 ) have already been related to the senescence in in vitro Bone Marrow 32 . ANKRD1 has already been related to the senescence in vivo of hMSC from bone marrow of older donors 46 , and IGFBP5 was upregulated in senescent cells 47 , 48 . In the list of 279 differentially expressed genes in the senescent hMSC/inv compared young cells, 22 ( AGTR1 , ANK2 , ANKRD 1 , BACE2 , DAB2 , HGF , JAM2 , LTBP2 , MOCOS , NMI , NRP1 , NTN4 , OXTR , PLXDC2 , SERPINB2 , SERPINE1 , SNCAIP , SPOCD1 , ST6GAL1 , SYNPO2 , THBS1 , TM4SF1 ) have also been identified as related to in vitro senescence 32 , and 10 genes ( RGS4 , ANKRD1 , NRXN3 , DDIT4 , C1R , PDE4DIP , GAS1 , CXCL12 , FST , C1S ) also has been related to aging in vivo 46 , and IGFP5 and SERPINE1 was associated to senescence cell from bone marrow and adipose tissue 47 .…”

Nuclear Nox4 interaction with prelamin A is associated with nuclear redox control of stem cell aging

“…In general, oxidative stress due to high ROS levels impairs stem cell homeostasis and can induce DNA damage, cell cycle arrest and eventually a senescence phenotype [ 5 ]. Indeed, long-term ASC expansion at low O 2 (5%) revoked in part the replicative senescence-associated alterations [ 6 ].…”

Secretome of Senescent Adipose-Derived Mesenchymal Stem Cells Negatively Regulates Angiogenesis

“…At early passages share of SA-β-gal-positive ASCs was very low. Cell size and granularity increase are considered senescence signs as well [23,28,29]. These changes are associated with the active metabolism, including synthetic processes, without cell division and with accumulation of non-utilizable lipofuscin granules.…”

“…Irreversible arrest of the cell cycle occurs, the morphology, organelles activity, and gene expression are altered, γH2AX heterochromatin foci appear, and a number of other cell senescence markers are found. Senescent cells are able to maintain their viability and functional activity for a rather long period, continuing to interact with the microenvironment and providing local and systemic effects [19][20][21][22][23].…”