Methods in molecular biology (Clifton, N.J.)Mar 7, 2019

Methods to Obtain, Identify, and Measure Aging in Human Fat-Derived Stem Cells

Longevity & Aging Weekly Brief ↗PubMed ↗DOI ↗

Updated Apr 4, 2026

Abstract

Human adipose-derived mesenchymal stem cells (hADSC) may experience replicative exhaustion and senescence during industrial-scale expansion.

Replicative senescence, characterized by the 'Hayflick limit,' significantly affects the in vivo effectiveness of stem cell cultures.
Senescence could contribute to clinical inconsistencies seen in infused cell preparations.
Characterization of hADSC senescence phenotypes is essential for understanding their function and therapeutic potential.
Protocols for isolating and culturing hADSC are provided, focusing on differentiation and characterization methods.
Key markers of cellular senescence, including immunophenotype and proliferation rate, are outlined for assessing hADSC quality.

Simplified

Human adipose-derived mesenchymal stem (stromal) cells (hADSC) represent an attractive source of the cells for numerous therapeutic applications in regenerative medicine. These cells are also an efficient model to study biological pathways of stem cell action, tissue injury and disease. Like any other primary somatic cells in culture, industrial-scale expansion of mesenchymal stromal cells (MSC) leads to the replicative exhaustion/senescence as defined by the "Hayflick limit." The senescence is not only greatly effecting in vivo potency of the stem cell cultures but also might be the cause and the source of clinical inconsistency arising from infused cell preparations. In this light, the characterization of hADSC replicative and stressor-induced senescence phenotypes is of great interest.This chapter summarizes some of the essential protocols and assays used at our laboratories and clinic for the human fat procurement, isolation, culture, differentiation, and characterization of mesenchymal stem cells from adipose tissue and the stromal vascular fraction. Additionally, we provide manuals for characterization of hADSC senescence in a culture based on stem cells immunophenotype, proliferation rate, migration potential, and numerous other well-accepted markers of cellular senescence. Such methodological framework will be immensely helpful to design standards and surrogate measures for hADSC-based therapeutic applications.

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Methods to Obtain, Identify, and Measure Aging in Human Fat-Derived Stem Cells

Abstract

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