Premium
Activation of rat nucleus pulposus cells by coculture with whole bone marrow cells collected by the perfusion method
Author(s) -
Umeda Masayuki,
Kushida Taketoshi,
Sasai Kunihiko,
Asada Taku,
Oe Kenichi,
Sakai Daisuke,
Mochida Joji,
Ikehara Susumu,
Iida Hirokazu
Publication year - 2009
Publication title -
journal of orthopaedic research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.041
H-Index - 155
eISSN - 1554-527X
pISSN - 0736-0266
DOI - 10.1002/jor.20740
Subject(s) - aggrecan , nucleus , bone marrow , mesenchymal stem cell , intervertebral disc , matrix (chemical analysis) , microbiology and biotechnology , perfusion , chemistry , immunology , pathology , medicine , anatomy , biology , alternative medicine , chromatography , osteoarthritis , articular cartilage
Cell proliferation and matrix synthesis were compared for rat nucleus pulposus cells cocultured with mesenchymal stem cells (MSCs) or fresh whole bone marrow cells (BMCs), harvested by the perfusion or aspiration methods. Nucleus pulposus cells were isolated from tail intervertebral discs of F344/slc rats, and BMCs were obtained from femora. Proteoglycan synthesis, DNA synthesis, and aggrecan mRNA expression were measured. The level of transforming growth factor‐β in supernatants from the culture system was also measured. Cell number, aggrecan mRNA expression, and uptake of [ 35 S]‐sulfate and [ 3 H]‐thymidine by nucleus pulposus cells cocultured with fresh whole BMCs all increased significantly compared with nucleus pulposus cells cocultured with MSCs. TGF‐β secreted by nucleus pulposus cells cocultured with fresh whole BMCs also significantly increased when compared with cocultures with MSCs. The perfusion method was superior to the aspiration method for preventing contamination of BMCs with peripheral red blood cells and lymphocytes, which may cause an autoimmune response in the disc. In conclusion, we suggest that fresh whole BMCs harvested by the perfusion method are more effective for increasing the proliferative and matrix synthesis capacity of nucleus pulposus cells. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:222–228, 2009