Tohoku J. Exp. Med., 2008, 216(2)

Differentiation of Adipose-derived Stromal Vascular Fraction Culture Cells into Chondrocytes Using the Method of Cell Sorting with a Mesenchymal Stem Cell Marker

DAISUKE ISHIMURA,¹ NAOKI YAMAMOTO,² KAORI TAJIMA,¹ AYUMI OHNO,¹ YASUHIRO YAMAMOTO,¹ OSUKE WASHIMI¹ and HARUMOTO YAMADA¹

¹Department of Orthopaedic Surgery, School of Medicine, Fujita Health University, Aichi, Japan
²Laboratory of Molecular Biology and Histochemistry, Joint Research Laboratory, Fujita Health University, Aichi, Japan

The incidence of arthritic diseases is rapidly increasing in most advanced countries. Articular cartilage, which is the most important tissue in the joint, consists of chondrocytes and abundant extracellular matrix, including aggrecan, and shows poor self-repair. We studied the potential of stem cells in mouse subcutaneous adipose tissue as a source of cells to regenerate cartilage tissue. Analysis of adipose-derived stromal vascular fraction culture cells (ADSVFs) using mesenchymal stem cell markers showed that CD90-positive cells accounted for 93.8%, CD105-positive cells for 68.5%, and p75 neurotrophin receptor (p75NTR, CD271)-positive cells for 36.1%. These results indicate that cells positive for mesenchymal stem cell markers are present in ADSVFs. The CD105-positive or -negative cells were isolated from ADSVFs by magnetic cell separation (MACS), and the efficiency of differentiation into chondrocytes was compared with using three methods of pellet method, gel-coating method, and gel-embedding sheet method. Using the CD105-positive cells and the gel-embedding sheet method, aggrecan mRNA was detected about three times higher than pellet and gel-coating methods. The above data suggest that ADSVFs could be differentiated into chondrocyte-like cells in the gel-embedding sheet method and could be useful in regenerative medicine to treat cartilage defects or cartilage degenerative disease. The use of cells sorted by mesenchymal stem cell markers from adipose tissue would gain position in the repair of cartilage tissue.

keywords —— chondrocyte; adipose-derived stromal vascular fraction culture cells (ADSVFs); CD105; mesenchymal stem cell; gel-embedding sheet.

© 2008 Tohoku University Medical Press

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Tohoku J. Exp. Med., 2008, 216, 149-156

Correspondence: Naoki Yamamoto, Laboratory of Molecular Biology and Histochemistry, Joint Research Laboratory, Fujita Health University, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake-city, Aichi 470-1192, Japan.

e-mail: naokiy@fujita-hu.ac.jp