Growth factors for clinical-scale expansion of human articular chondrocytes: relevance for automated bioreactor systems.

The expansion of chondrocytes in automated bioreactors for clinical use requires that a relevant number of cells be generated, starting from variable initial seeding densities in one passage and using autologous serum. We investigated whether the growth factor combination transforming growth factor beta 1/fibroblast growth factor 2/platelet-derived growth factor BB (TFP), recently shown to enhance the proliferation capacity of human articular chondrocytes (HACs), allows the efficiency of chondrocyte use to be increased at different seeding densities and percentages of human serum (HS). HACs were seeded at 1,000, 5,000, and 10,000 cells/cm2 in medium containing 10% fetal bovine serum or 10,000 cells/cm2 with 1%, 5%, or 10%HS. The chondrogenic capacity of post-expanded HACs was then assessed in pellet cultures. Expansion with TFP allowed a sufficient number of HACs to be obtained in one passage even at the lowest seeding density and HS percentage and variability in cartilage-forming capacity of HACs expanded under the different conditions to be reduced. Instead, larger variations and insufficient yields were found in the absence of TFP. By allowing large numbers of cells to be obtained, starting from a wide range of initial seeding densities and HS percentages, the use of TFP may represent a viable solution for the efficient expansion of HACs and addresses constraints of automated clinical bioreactor systems.

[1]  A I Caplan,et al.  In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. , 1998, Experimental cell research.

[2]  R. Sader,et al.  Experimental and mathematical study of the influence of growth factors on the growth kinetics of adult human articular chondrocytes , 2005, Journal of cellular physiology.

[3]  Mark H. Smith,et al.  Optimal in‐vitro expansion of chondroprogenitor cells in monolayer culture , 2006, Biotechnology and bioengineering.

[4]  J A Skinner,et al.  Autologous chondrocyte implantation versus matrix-induced autologous chondrocyte implantation for osteochondral defects of the knee: a prospective, randomised study. , 2005, The Journal of bone and joint surgery. British volume.

[5]  C. Brantsing,et al.  Human Serum for Culture of Articular Chondrocytes , 2005, Cell transplantation.

[6]  J. Verhaar,et al.  Multiplication of human chondrocytes with low seeding densities accelerates cell yield without losing redifferentiation capacity. , 2004, Tissue engineering.

[7]  P. Mainil-Varlet,et al.  A static, closed and scaffold-free bioreactor system that permits chondrogenesis in vitro. , 2003, Osteoarthritis and cartilage.

[8]  M. Heberer,et al.  Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro , 2001, Journal of cellular biochemistry.

[9]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[10]  M. Brittberg,et al.  Articular Cartilage Engineering with Autologous Chondrocyte Transplantation , 2003 .

[11]  Ivan Martin,et al.  Plasticity of clonal populations of dedifferentiated adult human articular chondrocytes. , 2003, Arthritis and rheumatism.

[12]  M. Brittberg,et al.  Articular cartilage engineering with autologous chondrocyte transplantation. A review of recent developments. , 2003, The Journal of bone and joint surgery. American volume.

[13]  Ivan Martin,et al.  Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity. , 2004, Osteoarthritis and cartilage.

[14]  M. Eloit Risks of virus transmission associated with animal sera or substitutes and methods of control. , 1999, Developments in biological standardization.

[15]  B. Oakes,et al.  The use of debrided human articular cartilage for autologous chondrocyte implantation: Maintenance of chondrocyte differentiation and proliferation in type I collagen gels , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  M. Brittberg,et al.  Two- to 9-Year Outcome After Autologous Chondrocyte Transplantation of the Knee , 2000, Clinical orthopaedics and related research.

[17]  B. Cahill,et al.  Weight training-related injuries in the high school athlete , 1982, The American journal of sports medicine.

[18]  C. Ohlsson,et al.  Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. , 1994, The New England journal of medicine.

[19]  Growth and Phenotype of Low-Density Nasal Septal Chondrocyte Monolayers , 2005, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[20]  C. Rorabeck,et al.  Increased damage to type II collagen in osteoarthritic articular cartilage detected by a new immunoassay. , 1994, The Journal of clinical investigation.

[21]  R. Sah,et al.  Human serum for tissue engineering of human nasal septal cartilage , 2006, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[22]  S. J. Wessman,et al.  Benefits and risks due to animal serum used in cell culture production. , 1999, Developments in biological standardization.

[23]  F. Grassi,et al.  Autologous Chondrocyte Implantation Using a Bilayer Collagen Membrane: A Preliminary Report , 2003, Journal of orthopaedic surgery.

[24]  F. Watt Effect of seeding density on stability of the differentiated phenotype of pig articular chondrocytes in culture. , 1988, Journal of cell science.

[25]  Maurilio Marcacci,et al.  Articular Cartilage Engineering with Hyalograft® C: 3-Year Clinical Results , 2005, Clinical orthopaedics and related research.

[26]  Maurilio Marcacci,et al.  Patellofemoral Full-Thickness Chondral Defects Treated with Hyalograft-C , 2006, The American journal of sports medicine.