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Methods and Strategies for Procurement, Isolation, Characterization, and Assessment of Senescence of Human Mesenchymal Stem Cells from Adipose Tissue

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Stem Cells and Aging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2045))

Abstract

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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Authors and Affiliations

  1. Aelan Cell Technologies, San Francisco, CA, USA

    Meenakshi Gaur & Victoria V. Lunyak

  2. Division of Plastic Surgery, University of California, San Diego, La Jolla, CA, USA

    Marek Dobke

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  1. Meenakshi Gaur
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  2. Marek Dobke
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  3. Victoria V. Lunyak
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Correspondence to Marek Dobke or Victoria V. Lunyak .

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  1. Ottawa, ON, Canada

    Kursad Turksen

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Gaur, M., Dobke, M., Lunyak, V.V. (2018). Methods and Strategies for Procurement, Isolation, Characterization, and Assessment of Senescence of Human Mesenchymal Stem Cells from Adipose Tissue. In: Turksen, K. (eds) Stem Cells and Aging . Methods in Molecular Biology, vol 2045. Humana, New York, NY. https://doi.org/10.1007/7651_2018_174

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