Tohoku J. Exp. Med., 2010, 221(2)

Protein Expression Profiles during Osteogenic Differentiation of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

SOLLIP KIM,¹ WON-KI MIN,¹ SAIL CHUN,¹ WOOCHANG LEE,¹ HEE-JUNG CHUNG,¹ SOO JIN CHOI,² SUNG EUN YANG,² YOON SUN YANG² and JONG-IL YOO³

¹Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea
²Department of Research and Development for Cellular Therapy, Medipost Biomedical Research Institute, Seoul, Korea
³In2Gen Co., Ltd., Seoul, Korea

Mesenchymal stem cells (MSCs) can potentially differentiate along multiple lineages and be expanded in vitro, making them highly attractive candidates for cell therapy and tissue engineering applications. This study sought to investigate the critical proteins involved in osteogenic differentiation of mesenchymal stem cells derived from umbilical cord blood (UCB-MSCs). MSCs, which were isolated from three different preparations of human UCB, were osteoinduced, and total proteins were extracted from the cells. Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was performed on the day (d) of induction d0, and on d2, d7, and d21 of differentiation. The optical density (OD) of each spot was measured, and spots with a mean OD of three cell lines of MSCs that increased > 30 or decreased < 0.1 relative to a previous time point were selected. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF/MS) was used to identify the proteins. Through database searches, the properties and functions of the proteins were investigated and then classified according to the Gene Ontology classification. Among the 308 spots observed in the 2-D gel, 16 proteins with a mean OD ratio > 30, and 20 proteins with a mean OD ratio < 0.1 were identified during the differentiation process. Additionally, the distribution of differentially expressed proteins according to cellular component and molecular function criteria differed depending on whether protein expression increased or decreased during differentiation. The results of this study will comprise an initial proteomic database for UCB-MSCs differentiation.

keywords —— differentiation; mesenchymal stem cell; osteogenic; proteomics; umbilical cord blood

© 2010 Tohoku University Medical Press

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Tohoku J. Exp. Med., 2010, 221, 141-150

Correspondence: Won-Ki Min, M.D., Ph.D., Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, 388-1 Pungnap-dong, Songpa-Gu, Seoul, 138-736, Korea.

e-mail: WKmin@amc.seoul.kr