Live-Cell, Temporal Gene Expression Analysis of Osteogenic Differentiation in Adipose-Derived Stem Cells
作者： Hetal V. Desai , Indu S. Voruganti , Chathuraka Jayasuriya , Qian Chen , Eric M. Darling
DOI: 10.1089/TEN.TEA.2012.0127
关键词: Gene expression profiling 、 Spatiotemporal gene expression 、 Regulation of gene expression 、 Musculoskeletal regeneration 、 Biology 、 Molecular biology 、 Cellular differentiation 、 Stem cell 、 ALPL 、 Population
摘要: Adipose-derived stem cells (ASCs) are a widely investigated type of mesenchymal cell with great potential for musculoskeletal regeneration. However, use ASCs is complicated by their cellular heterogeneity, which exists at both the population and single-cell levels. This study demonstrates live-cell assay to investigate gene expression in undergoing osteogenesis using fluorescently tagged DNA hybridization probes called molecular beacons. Three beacons were designed target mRNA sequences alkaline phosphatase, I collagen, osteocalcin (ALPL, COL1A1, BGLAP), genes characteristically expressed during osteogenesis. The percentage expressing these was monitored daily quantify uniformity differentiation process. Differentiating ASC populations repeatedly measured nondestructive fashion over 21-day period obtain temporal data. Results showed consistent patterns osteogenic response induction medium. Peak observed days 3-4 ALPL, day 14 21 BGLAP. Additionally, sample became more uniform after 2 weeks medium, suggesting syncing occurs time. These findings previous studies suggest that viable means monitor spatiotemporal live, differentiating ASCs.
sci-hub.se 参考文章(40)
Diana P. Bratu, Irina E. Catrina, Salvatore A. E. Marras, Tiny Molecular Beacons for in vivo mRNA Detection Methods in Molecular Biology. ,vol. 714, pp. 141- 157 ,(2011) , 10.1007/978-1-61779-005-8_9
David H. Mathews, Douglas H. Turner, Michael Zuker, RNA secondary structure prediction. Current protocols in human genetics. ,vol. 67, pp. 112- ,(2000) , 10.1002/0471142700.NC1102S28
Jeffrey M Gimble, Warren Grayson, Farshid Guilak, Mandi J Lopez, Gordana Vunjak-Novakovic, Adipose tissue as a stem cell source for musculoskeletal regeneration Frontiers in Bioscience. ,vol. S3, pp. 69- 81 ,(2011) , 10.2741/S133
Hermann Agis, Stefan Schröckmair, Carmine Skorianz, Michael B. Fischer, Georg Watzek, Reinhard Gruber, Platelets increase while serum reduces the differentiation and activity of osteoclasts in vitro Journal of Orthopaedic Research. ,vol. 31, pp. 1561- 1569 ,(2013) , 10.1002/JOR.22386
Jeffrey M. Gimble, Farshid Guilak, Mark E. Nuttall, Solomon Sathishkumar, Martin Vidal, Bruce A. Bunnell, In vitro Differentiation Potential of Mesenchymal Stem Cells. Transfusion Medicine and Hemotherapy. ,vol. 35, pp. 228- 238 ,(2008) , 10.1159/000124281
Bradley T. Estes, Arthur W. Wu, Robert W. Storms, Farshid Guilak, Extended passaging, but not aldehyde dehydrogenase activity, increases the chondrogenic potential of human adipose-derived adult stem cells. Journal of Cellular Physiology. ,vol. 209, pp. 987- 995 ,(2006) , 10.1002/JCP.20808
David W. Mount, Using the Basic Local Alignment Search Tool (BLAST) CSH Protocols. ,vol. 2007, ,(2007) , 10.1101/PDB.TOP17
Katie C. Russell, Donald G. Phinney, Michelle R. Lacey, Bonnie L. Barrilleaux, Kristin E. Meyertholen, Kim C. O'Connor, In Vitro High-Capacity Assay to Quantify the Clonal Heterogeneity in Trilineage Potential of Mesenchymal Stem Cells Reveals a Complex Hierarchy of Lineage Commitment Stem Cells. ,vol. 28, pp. 788- 798 ,(2010) , 10.1002/STEM.312
David E. Rothem, Lilah Rothem, Michael Soudry, Aviva Dahan, Rami Eliakim, Nicotine modulates bone metabolism-associated gene expression in osteoblast cells Journal of Bone and Mineral Metabolism. ,vol. 27, pp. 555- 561 ,(2009) , 10.1007/S00774-009-0075-5
Eung-Kyun Kim, Seyoung Lim, Ji-Man Park, Jeong Kon Seo, Jae Ho Kim, Kyong Tai Kim, Sung Ho Ryu, Pann-Ghill Suh, Human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by AMP‐activated protein kinase Journal of Cellular Physiology. ,vol. 227, pp. 1680- 1687 ,(2012) , 10.1002/JCP.22892