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After long-term culture, mesenchymal stem cells alter their biological properties and enter into a state of replicative senescence. Although several classical biomarkers have been used for quantitative assessment of cellular senescence, no hallmark has been proven completely unique to the senescent state in cells. We used bone marrow-derived MSCs (BM-MSCs) from different healthy young donors and an  in vitro  model with well-defined senescence end points to identify a set of robust markers that could potentially predict the expansion capacity of MSCs preparations before reaching senescence. For each early passage BM-MSC sample (5th or 6th passages), the normalized protein expression levels of senescence-associated markers p16 INK4A , p21 WAF1 , SOD2, and rpS6 S240/244 ; the concentration of IL6 and IL8 in cell culture supernatants; and the normalized gene expression levels of pluripotency markers  OCT4 ,  NANOG , and  SOX2  were correlated with final population doubling (PD) number. We revealed that the low expression of p16 INK4A  protein and a high  OCT4  gene expression, rather than other evaluated markers, might be potential hallmarks and predictors of greater  in vitro  lifespan and growth potential, factors that can impact the successful therapeutic use of MSCs preparations.

stem cells international

High<i>OCT4</i>and Low<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M1"><mml:mrow><mml:msup><mml:mrow><mml:mtext>p16</mml:mtext></mml:mrow><mml:mrow><mml:mtext>INK4A</mml:mtext></mml:mrow></mml:msup></mml:mrow></mml:math>Expressions Determine<i>In Vitro</i>Lifespan of Mesenchymal Stem Cells

HighOCT4and Lowp16INK4AExpressions DetermineIn VitroLifespan of Mesenchymal Stem Cells