Establishment of In Vitro Models by Stress-Induced Premature Senescence for Characterizing the Stromal Vascular Niche in Human Adipose Tissue

被引：2

作者：

Wahlmueller, Marlene ^{[1
,2
,3
]}

Narzt, Marie-Sophie ^{[1
,2
,3
]}

Missfeldt, Karin ^{[1
,2
]}

Arminger, Verena ^{[1
,2
]}

Krasensky, Anna ^{[1
,2
]}

Laemmermann, Ingo ^{[4
,5
]}

Schaedl, Barbara ^{[1
,2
,6
]}

Mairhofer, Mario ^{[7
]}

Suessner, Susanne ^{[8
]}

Wolbank, Susanne ^{[1
,2
]}

Priglinger, Eleni ^{[1
,2
,3
]}

机构：

[1] Ludwig Boltzmann Inst Traumatol Cooperat AUVA, A-1200 Vienna, Austria

[2] Austrian Cluster Tissue Regenerat, A-1200 Vienna, Austria

[3] MorphoMed GmbH, A-1030 Vienna, Austria

[4] Univ Nat Resources & Life Sci, Inst Mol Biotechnol, Dept Biotechnol, Christian Doppler Lab Biotechnol Skin Aging, A-1190 Vienna, Austria

[5] Rockfish Bio AG, A-1010 Vienna, Austria

[6] Med Univ Vienna, Univ Clin Dent, A-1090 Vienna, Austria

[7] Johannes Kepler Univ Linz, Dept Hematol & Internal Oncol, A-4020 Linz, Austria

[8] Austrian Red Cross Blood Transfus Serv Upper Aust, A-4020 Linz, Austria

来源：

LIFE-BASEL | 2022年 / 12卷 / 10期

关键词：

senescence; stress-induced premature senescence (SIPS); doxorubicin; adipose tissue; adipose-derived stromal; stem cells (ASCs); stromal vascular fraction (SVF); microtissue; CELLULAR SENESCENCE; CELLS; DOXORUBICIN; ARREST; INHIBITION; TRANSITION; INDUCTION; BIOMARKER; CULTURE; GROWTH;

D O I：

10.3390/life12101459

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Acting as the largest energy reservoir in the body, adipose tissue is involved in longevity and progression of age-related metabolic dysfunction. Here, cellular senescence plays a central role in the generation of a pro-inflammatory environment and in the evolution of chronic diseases. Within the complexity of a tissue, identification and targeting of senescent cells is hampered by their heterogeneity. In this study, we generated stress-induced premature senescence 2D and 3D in vitro models for the stromal vascular niche of human adipose tissue. We established treatment conditions for senescence induction using Doxorubicin (Dox), starting from adipose-derived stromal/stem cells (ASCs), which we adapted to freshly isolated microtissue-stromal vascular fraction (MT-SVF), where cells are embedded within their native extracellular matrix. Senescence hallmarks for the established in vitro models were verified on different cellular levels, including morphology, cell cycle arrest, senescence-associated beta-galactosidase activity (SA-beta gal) and gene expression. Two subsequent exposures with 200 nM Dox for six days were suitable to induce senescence in our in vitro models. We demonstrated induction of senescence in the 2D in vitro models through SA-beta gal activity, at the mRNA level (LMNB1, CDK1, p21) and additionally by G2/M phase cell cycle arrest in ASCs. Significant differences in Lamin B1 and p21 protein expression confirmed senescence in our MT-SVF 3D model. MT-SVF 3D cultures were composed of multiple cell types, including CD31, CD34 and CD68 positive cells, while cell death remained unaltered upon senescence induction. As heterogeneity and complexity of adipose tissue senescence is given by multiple cell types, our established senescence models that represent the perivascular niche embedded within its native extracellular matrix are highly relevant for future clinical studies.

引用

页数：20

共 55 条

[1] Adipose-derived stromal vascular fraction cells isolated from old animals exhibit reduced capacity to support the formation of microvascular networks [J].