Downregulation of Melanoma Cell Adhesion Molecule (MCAM/CD146) Accelerates Cellular Senescence in Human Umbilical Cord Blood‐Derived Mesenchymal Stem Cells

• Published in: Stem cells translational medicine 2016-04, Vol.5 (4), p.427-439
• Main Authors: Jin, Hye Jin, Kwon, Ji Hye, Kim, Miyeon, Bae, Yun Kyung, Choi, Soo Jin, Oh, Wonil, Yang, Yoon Sun, Jeon, Hong Bae
• Format: Article
• Language: English
• Subjects: Biomarkers; Biomarkers - metabolism; Bmi‐1; CD146 Antigen - genetics; CD146 Antigen - metabolism; Cell adhesion & migration; Cell adhesion molecules; Cell culture; Cell cycle; Cell Membrane - drug effects; Cell Membrane - metabolism; Cell Proliferation - drug effects; Cell Proliferation - genetics; Cell surface marker; Cellular senescence; Cellular Senescence - drug effects; Cellular Senescence - genetics; Cord blood; Data analysis; Down-Regulation; Fetal and Neonatal Stem Cells; Fetal Blood - cytology; Fetal Blood - metabolism; Flow cytometry; Gene Expression Regulation - drug effects; Human umbilical cord blood-derived mesenchymal stem cells; Humans; Id1; Id1 protein; Immunoglobulins; Infant, Newborn; Melanoma; Melanoma cell adhesion molecule (CD146); Mesenchymal Stem Cell Transplantation - standards; Mesenchymal stem cell-based therapy; Mesenchymal stem cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - drug effects; Mesenchymal Stromal Cells - metabolism; Mesenchyme; Morphology; Phosphorylation; Proteins; Quality Control; RNA, Small Interfering - pharmacology; Senescence; Stem cells; Studies; Transcription factors; Twist1; Umbilical cord
• ISSN: 2157-6564; 2157-6580
• DOI: 10.5966/sctm.2015-0109

CD146 expression is gradually decreased during the long‐term expansion of human umbilical cord blood‐derived mesenchymal stem cells (hUCB‐MSCs) in culture, and enforced CD146 downregulation accelerated senescence in hUCB‐MSCs, which affected their multilineage differentiation potential, growth, and stemness. These data indicate a possible role for CD146 in inhibiting senescence in hUCB‐MSCs, which may occur via the regulation of Bmi‐1, Id1, and Twist1 expression. CD146 may be a novel marker for predicting senescence in hUCB‐MSCs, and it could be valuable in quality‐control assessments and for improving the therapeutic potential of hUCB‐MSC‐based therapy. Therapeutic applications of mesenchymal stem cells (MSCs) for treating various diseases have increased in recent years. To ensure that treatment is effective, an adequate MSC dosage should be determined before these cells are used for therapeutic purposes. To obtain a sufficient number of cells for therapeutic applications, MSCs must be expanded in long‐term cell culture, which inevitably triggers cellular senescence. In this study, we investigated the surface markers of human umbilical cord blood‐derived MSCs (hUCB‐MSCs) associated with cellular senescence using fluorescence‐activated cell sorting analysis and 242 cell surface‐marker antibodies. Among these surface proteins, we selected the melanoma cell adhesion molecule (MCAM/CD146) for further study with the aim of validating observed expression differences and investigating the associated implications in hUCB‐MSCs during cellular senescence. We observed that CD146 expression markedly decreased in hUCB‐MSCs following prolonged in vitro expansion. Using preparative sorting, we found that hUCB‐MSCs with high CD146 expression displayed high growth rates, multilineage differentiation, expression of stemness markers, and telomerase activity, as well as significantly lower expression of the senescence markers p16, p21, p53, and senescence‐associated β‐galactosidase, compared with that observed in hUCB‐MSCs with low‐level CD146 expression. In contrast, CD146 downregulation with small interfering RNAs enhanced the senescence phenotype. In addition, CD146 suppression in hUCB‐MSCs caused downregulation of other cellular senescence regulators, including Bmi‐1, Id1, and Twist1. Collectively, our results suggest that CD146 regulates cellular senescence; thus, it could be used as a therapeutic marker to identify senescent hUCB‐MSCs. Significance One of the fundament