title: Isolation, culture, and osteogenic/chondrogenic differentiation of bone marrow-derived mesenchymal stem cells.
abstract: Musculoskeletal disorders, as non-healing fractures and large bone defects, articular cartilage and subchondral bone injuries, often result in lifelong chronic pain and compromised quality of life. Although generally a natural process, failure of large bone defects to heal such as after complex fractures, resection of tumours, infections, or revisions of joint replacements remains a critical challenge that requires more appropriate solutions as those currently available. In addition, regeneration of chondral and osteochondral defects continues to be a challenge until to date. A profound understanding of the underlying mechanisms of endogenous regeneration is a prerequisite for successful bone and cartilage regeneration. Presently, one of the most promising therapeutic approaches is cell-based tissue engineering which provides a healthy population of cells to the injured site. Use of differentiated cells has severe limitations; an excellent alternative would be the application of adult marrow stromal cells/mesenchymal stem cells (MSC) which possess extensive proliferation potential and proven capability to differentiate along the osteochondral pathway. The process of osteo-/chondrogenesis can be mimicked in vitro by inducing osteo-chondroprogenitor stem cells to undergo osteogenesis and chondrogenesis through exposure of osteo-/chondrogenic favourable microenvironmental, mechanical, and nutritional conditions. This chapter provides comprehensive protocols for the isolation, expansion, and osteo-/chondrogenic differentiation of adult bone marrow-derived MSC.
authors.affiliation: Department of Orthopaedics, University of Regensburg and Centre for Biomedical Technology, BioPark, Regensburg, Germany. susanne.graessel@klinik.uni-regensburg.de
authors.forename: Susanne
authors.initials: S
authors.lastname: Grässel
authors.forename: Sabine
authors.initials: S
authors.lastname: Stöckl
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Jenei-Lanzl
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002417:Cattle; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002645:Chickens; D020219:Chondrogenesis; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D010012:Osteogenesis; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013552:Swine