IJCEM Copyright © 2008-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711
Int J Clin Exp Med 2012;5(1):34-43

Original Article
Establishment of rat bone mesenchymal stem cell lines stably expressing
Chondromodulin I

Shuangchun Xing*, Zhiqiang Wang*, Hui Xi, Lianzhong Zhou, Dongqing Wang, Lin Sang, Xinhui Wang, Min Qi, Lijie Zhai

Department of Otolaryngology-Head and Neck Surgery, First affiliated hospital of Dalian Medical University, Dalian 116011, P.R.China;
Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, P.R.China; Department of Cardiology, Beijing Anzhen Hospital,
Capital Medical University, Beijing 100029, P.R.China; School of Materials Science and Engineering, Dalian University of Technology,
Dalian 116024, P R China. *Equal contributors.

Received November 29, 2011; accepted January 12, 2012; Epub January 15, 2012; Published January 31, 2012

Abstract: To study the role of Chondromodulin I (ChM-I) in inducing bone mesenchymal stem cells (MSCs) into chondrocytes, the
plasmid expressing pcDNA3.1 (+) / ChM-I was constructed, and transfected into MSCs. Stable expression verified that ChM-I is
upregulated in MSCs, which make it a useful preparation for studying the further role of ChM-I in inducing BMSCs toward a
chondrogenic I phenotype and for tissue engineering constructs. MSCs were obtained from adult Sprague Dawley (SD) rats using
density gradient centrifugation. Cell surface antigens were detected by flow cytometry. The experimental groups were MSCs transfected
with the plasmid pcDNA3.1 (+)/ChM-I. The control one was with the plasmid pcDNA3.1 (+) and the negative control one was with no
plasmid. Transcription and translation were detected in the level of mRNA and protein respectively in order to identify the stability of the
expression of ChM-I in cell lines by RT-PCR and western blot analysis. The results of RT-PCR showed that the mRNA level of ChM-I in
the experimental group was six times higher than the control. The quantitative optical density analysis showed that the expression of
ChM-I in the experimental group was significantly higher than the control by western blot. We conclude that we have established MSCs
lines stably expressing ChM-I. This work lays the foundation of studying the potential role of the ChM-I in inducing BMSCs into
chondrocytes.
(IJCEM1111009).

Keywords: Chondromodulin-I, MSCs biological factors, plasmid pcDNA3.1 (+)/ChM-I, tissue engineering

Address all correspondence to:
Dr. Li-Jie Zhai
Department of Otolaryngology-Head and Neck Surgery
First affiliated hospital of Dalian Medical University
222 Zhongshan RD, Dalian, China 116011.
Tel: 86-411-83635963; Fax: 86-411-83673291
E-mail: lijiezhai@yahoo.com.cn