In vitro cartilage formation using TGF-beta-immobilized magnetic beads and mesenchymal stem cell-magnetic bead complexes under magnetic field conditions.
We evaluated the efficacy of transforming growth factor (TGF)-beta-immobilized magnetic beads for chondrogenesis in vitro using a mesenchymal stem cell (MSC) delivery system and an external magnetic force (EMF). MSCs isolated from the bone marrow of Sprague Dawley rats were mixed with carboxyl group-combined magnetic beads (Ferri Sphere 100C) coated with anti-rat CD44 mouse monoclonal antibodies. TGF-beta3 (10 and 1 ng/mL) was attached magnetically to such other Ferri Sphere 100C beads via an amide bond formed between a primary amino group on the TGF-beta3 and the carboxyl groups on the surface of the beads. MSC-magnetic bead complexes were centrifuged to form a pellet and cultured in chondrogenic differentiation medium (CDM) supplemented with either 10 or 1 ng/mL TGF-beta-immobilized magnetic beads (10 or 1 ng/mL TGF-beta-immobilized magnetic bead groups) or in CDM supplemented with 1 or 10 ng/mL TGF-beta (1 or 10 ng/mL TGF-beta group). TGF-beta-immobilized magnetic beads were gathered effectively under an EMF. Chondrogenesis was achieved from the MSC-magnetic bead complexes in the presence of 1 ng/mL TGF-beta-immobilized magnetic beads.
Motoyama
Mitsuru
M
Department of Orthopaedics Surgery, Graduate School of Biomedical Sciences Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan. mmituru32@yahoo.co.jp
Deie
Masataka
M
Kanaya
Atsushi
A
Nishimori
Makoto
M
Miyamoto
Ayato
A
Yanada
Shinobu
S
Adachi
Nobuo
N
Ochi
Mitsuo
M
D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D020219:Chondrogenesis / Q000187:drug effects*; D018960:Hyaluronan Receptors / Q000378:metabolism; D055767:Immobilized Proteins / Q000494:pharmacology*; D008280:Magnetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects*; D008863:Microspheres; D009354:Neodymium; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology*
D018960:Hyaluronan Receptors; D055767:Immobilized Proteins; D053782:Transforming Growth Factor beta3; D009354:Neodymium