Original Article
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World J Stem Cells. Aug 26, 2011; 3(8): 70-82
Published online Aug 26, 2011. doi: 10.4252/wjsc.v3.i8.70
Characterization of mouse clonal mesenchymal stem cell lines established by subfractionation culturing method
Myung-Shin Jeon, Tac-Ghee Yi, Hyun-Ja Lim, Sun-Hwa Moon, Moon-Hee Lee, Joon-Soon Kang, Chul-Soo Kim, Dae-Hyun Lee, Sun U Song
Myung-Shin Jeon, Tac-Ghee Yi, Hyun-Ja Lim, Sun-Hwa Moon, Moon-Hee Lee, Joon-Soon Kang, Chul-Soo Kim, Dae-Hyun Lee, Sun U Song, Clinical Research Center, School of Medicine, Inha University, 7-206, 3-Ga, Shinheung-Dong, Chung-Gu, Inchon 400-711, South Korea
Author contributions: Jeon MS and Yi TG performed the majority of experiments; Lim HJ, Moon SH and Lee DH performed some experiments; Lee MH, Kang JS and Kim CS provided the materials and financial support for this work; Song SU designed the study and wrote the manuscript.
Supported by A Grant from the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea (A092142), a research grant from HomeoTherapy Co. Ltd. (39856-01) and by the Brain Korea 21 Project in 2010
Correspondence to: Dr. Sun U Song, PhD, Professor, Clinical Research Center, College of Medicine, Inha University, 7-206, 3-Ga, Shinheung-Dong, Chung-Gu, Inchon 400-711, South Korea. sunuksong@inha.ac.kr
Telephone: +82-32-8902460 Fax: +82-32-8902462
Received: August 10, 2010
Revised: January 10, 2011
Accepted: January 20, 2011
Published online: August 26, 2011
Abstract

AIM: To characterize single-cell-derived mouse clonal mesenchymal stem cells (mcMSCs) established with bone marrow samples from three different mouse strains.

METHODS: We established mcMSC lines using subfractionation culturing method from bone marrow samples obtained from long bones. These lines were characterized by measuring cell growth, cell surface epitopes, differentiation potential, lineage-specific gene expression and T-cell suppression capability. Nonclonal MSCs isolated by the conventional gradient centrifugation method were used as controls.

RESULTS: All mcMSC lines showed typical nonclonal MSC-like spindle shape morphology. Lines differed in optimal growth density requirement. Cell surface epitope profiles of these mcMSC lines were similar to those of nonclonal MSCs. However, some lines exhibited different expression levels in a few epitopes, such as CD44 and CD105. Differentiation assays showed that 90% of the mcMSC lines were capable of differentiating into adipogenic and/or chondrogenic lineages, but only 20% showed osteogenic lineage differentiation. T-cell suppression analysis showed that 75% of the lines exhibited T-cell suppression capability.

CONCLUSION: mcMSC lines have similar cell morphology and cell growth rate but exhibit variations in their cell surface epitopes, differentiation potential, lineage-specific gene expression and T-cell suppression capability.

Keywords: Clonal stem cells; Bone marrow stem cells; Multilineage differentiation; Clonal stromal cell isolation; T-cell suppression