间充质干细胞
干细胞
衰老
生物
细胞生物学
间质细胞
脂肪组织
基质血管部分
再生医学
间充质干细胞的临床应用
免疫学
细胞分化
癌症研究
成体干细胞
内分泌学
遗传学
基因
作者
Meenakshi Gaur,Marek Dobke,Victoria V. Lunyak
出处
期刊:Methods in molecular biology
日期:2018-01-01
卷期号:: 37-92
被引量:12
标识
DOI:10.1007/7651_2018_174
摘要
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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