Influence of species and anatomical location on chondrocyte expansion.
Abstract: Bovine articular cartilage is often used to study chondrocytes in vitro. It is difficult to correlate in vitro studies using bovine chondrocytes with in vivo studies using other species such as rabbits and sheep. The aim of this investigation was to study the effect of species, anatomical location and exogenous growth factors on chondrocyte proliferation in vitro. Methods: Equine (EQ), bovine (BO) and ovine (OV) articular chondrocytes from metacarpophalangeal (fetlock (F)), shoulder (S) and knee (K) joints were cultured in tissue culture flasks. Growth factors (rh-FGFb: 10 ng/ml; rh-TGFbeta: 5 ng/ml) were added to the cultures at days 2 and 4. On day 6, cells were counted and flow cytometry analysis was performed to determine cell size and granularity. A three factor ANOVA with paired Tukey's correction was used for statistical analysis. Results: After 6 days in culture, cell numbers had increased in control groups of EQ-F, OV-S, OV-F and BO-F chondrocytes. The addition of rh-FGFb led to the highest increase in cell numbers in the BO-F, followed by EQ-F and OV-S chondrocytes. The addition of rh-TGFbeta increased cell numbers in EQ-S and EQ-F chondrocytes, but showed nearly no effect on EQ-K, OV-K, OV-S, OV-F and BO-F chondrocytes. There was an overall difference with the addition of growth factors between the different species and joints. Conclusions: Different proliferation profiles of chondrocytes from the various joints were found. Therefore, we recommend performing in vitro studies using the species and site where subsequent in vivo studies are planned.
Publication Date: 2005-05-17 PubMed ID: 15904515PubMed Central: PMC1166560DOI: 10.1186/1471-2474-6-23Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study investigated the impact of species, anatomical location and added growth factors on the proliferation and growth of chondrocytes (cartilage cells). The research showed varying growth profiles for chondrocytes from different joints and species, and recommended performing experiments in species and sites where subsequent in-depth studies are planned.
Research Methodology
- The research employed equine (EQ), bovine (BO), and ovine (OV) articular chondrocytes from the metacarpophalangeal (fetlock (F)), shoulder (S), and knee (K) joints.
- These cells were cultured in tissue culture flasks, and certain growth factors (rh-FGFb: 10 ng/ml; rh-TGFbeta: 5 ng/ml) were added to the cultures at the second and fourth days.
- Cell count was done on the sixth day, and flow cytometry analysis was conducted to ascertain cell size and granularity.
- For statistical analysis, the team utilized a three-factor ANOVA with a paired Tukey’s correction.
Research Findings
- After six days in culture, an increase in cell numbers was noticed in control groups of EQ-F, OV-S, OV-F, and BO-F chondrocytes.
- The addition of rh-FGFb caused the highest increase in cell numbers in the BO-F, followed by EQ-F and OV-S chondrocytes.
- rh-TGFbeta increased cell numbers in EQ-S and EQ-F chondrocytes, but exhibited nearly no effect on EQ-K, OV-K, OV-S, OV-F, and BO-F chondrocytes.
- A noticeable difference with the addition of growth factors between the different species and joints was observed.
Conclusion and Recommendations
- The study concluded with different proliferation profiles of chondrocytes from the various joints.
- As such, it recommends performing in vitro studies using the species and site where subsequent in vivo studies are planned to maintain consistency and evaluate more accurately the cartilage cells’ behavior in a similar environment.
Cite This Article
APA
Akens MK, Hurtig MB.
(2005).
Influence of species and anatomical location on chondrocyte expansion.
BMC Musculoskelet Disord, 6, 23.
https://doi.org/10.1186/1471-2474-6-23 Publication
Researcher Affiliations
- Dept. of Clinical Studies, University of Guelph, Guelph, Ontario, N1G 2W1, Canada. makens@uhnres.utoronto.ca
MeSH Terms
- Animals
- Cartilage, Articular / cytology
- Cattle
- Cell Count
- Cell Division / drug effects
- Cell Proliferation / drug effects
- Cell Size
- Cells, Cultured
- Chondrocytes / cytology
- Chondrocytes / ultrastructure
- Cytoplasmic Granules / ultrastructure
- Fibroblast Growth Factor 2 / pharmacology
- Horses
- Humans
- Knee Joint
- Metacarpophalangeal Joint
- Recombinant Proteins / pharmacology
- Sheep
- Shoulder Joint
- Species Specificity
- Transforming Growth Factor beta / pharmacology
References
This article includes 49 references
- Lee DA, Bentley G, Archer CW. The control of cell division in articular chondrocytes.. Osteoarthritis Cartilage 1993 Apr;1(2):137-46.
- Glansbeek HL, van der Kraan PM, Lafeber FP, Vitters EL, van den Berg WB. Species-specific expression of type II TGF-beta receptor isoforms by articular chondrocytes: effect of proteoglycan depletion and aging.. Cytokine 1997 May;9(5):347-51.
- Martin I, Vunjak-Novakovic G, Yang J, Langer R, Freed LE. Mammalian chondrocytes expanded in the presence of fibroblast growth factor 2 maintain the ability to differentiate and regenerate three-dimensional cartilaginous tissue.. Exp Cell Res 1999 Dec 15;253(2):681-8.
- Cuevas P, Burgos J, Baird A. Basic fibroblast growth factor (FGF) promotes cartilage repair in vivo.. Biochem Biophys Res Commun 1988 Oct 31;156(2):611-8.
- Mow VC, Ratcliffe A, Rosenwasser MP, Buckwalter JA. Experimental studies on repair of large osteochondral defects at a high weight bearing area of the knee joint: a tissue engineering study.. J Biomech Eng 1991 May;113(2):198-207.
- Calvo E, Palacios I, Delgado E, Ruiz-Cabello J, Hernández P, Sánchez-Pernaute O, Egido J, Herrero-Beaumont G. High-resolution MRI detects cartilage swelling at the early stages of experimental osteoarthritis.. Osteoarthritis Cartilage 2001 Jul;9(5):463-72.
- Frenkel SR, Saadeh PB, Mehrara BJ, Chin GS, Steinbrech DS, Brent B, Gittes GK, Longaker MT. Transforming growth factor beta superfamily members: role in cartilage modeling.. Plast Reconstr Surg 2000 Mar;105(3):980-90.
- Weisser J, Rahfoth B, Timmermann A, Aigner T, Bräuer R, von der Mark K. Role of growth factors in rabbit articular cartilage repair by chondrocytes in agarose.. Osteoarthritis Cartilage 2001;9 Suppl A:S48-54.
- Tan CF, Ng KK, Ng SH, Cheung YC. Magnetic resonance imaging of hyaline cartilage regeneration in neocartilage graft implantation.. Transplant Proc 2003 Dec;35(8):3105-7.
- Hansen AL, Foster BK, Gibson GJ, Binns GF, Wiebkin OW, Hopwood JJ. Growth-plate chondrocyte cultures for reimplantation into growth-plate defects in sheep. Characterization of cultures.. Clin Orthop Relat Res 1990 Jul;(256):286-98.
- Schachar NS, Novak K, Hurtig M, Muldrew K, McPherson R, Wohl G, Zernicke RF, McGann LE. Transplantation of cryopreserved osteochondral Dowel allografts for repair of focal articular defects in an ovine model.. J Orthop Res 1999 Nov;17(6):909-19.
- Akens MK, von Rechenberg B, Bittmann P, Nadler D, Zlinszky K, Auer JA. Long term in-vivo studies of a photo-oxidized bovine osteochondral transplant in sheep.. BMC Musculoskelet Disord 2001;2:9.
- Jelic M, Pecina M, Haspl M, Kos J, Taylor K, Maticic D, McCartney J, Yin S, Rueger D, Vukicevic S. Regeneration of articular cartilage chondral defects by osteogenic protein-1 (bone morphogenetic protein-7) in sheep.. Growth Factors 2001;19(2):101-13.
- Muldrew K, Chung M, Novak K, Schachar NS, Zernicke RF, McGann LE, Rattner JB, Matyas JR. Evidence of chondrocyte repopulation in adult ovine articular cartilage following cryoinjury and long-term transplantation.. Osteoarthritis Cartilage 2001 Jul;9(5):432-9.
- Mohr W. Gelenkerkrankheiten : Diagnostik und Pathogenese makroskopischer und histologischer Strukturveränderungen. Stuttgart, Georg Thieme Verlag; 1984.
- Dieppe P, Kirwan J. The localization of osteoarthritis.. Br J Rheumatol 1994 Mar;33(3):201-3.
- Smith MM, Little CB, Rodgers K, Ghosh P. [Animal models used for the evaluation of anti-osteoarthritis drugs].. Pathol Biol (Paris) 1997 Apr;45(4):313-20.
- Lozada CJ, Altman RD. Animals Models of Cartilage Breakdown. Bone and Cartilage Metabolism San Diego, Academic Press; 1999. pp. 339–352.
- Shapiro F, Koide S, Glimcher MJ. Cell origin and differentiation in the repair of full-thickness defects of articular cartilage.. J Bone Joint Surg Am 1993 Apr;75(4):532-53.
- Hunziker EB. Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects.. Osteoarthritis Cartilage 2002 Jun;10(6):432-63.
- Armstrong SJ, Read RA, Price R. Topographical variation within the articular cartilage and subchondral bone of the normal ovine knee joint: a histological approach.. Osteoarthritis Cartilage 1995 Mar;3(1):25-33.
- Solchaga LA, Goldberg VM, Caplan AI. Cartilage regeneration using principles of tissue engineering.. Clin Orthop Relat Res 2001 Oct;(391 Suppl):S161-70.
- Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L. Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation.. N Engl J Med 1994 Oct 6;331(14):889-95.
- de Haart M, Marijnissen WJ, van Osch GJ, Verhaar JA. Optimization of chondrocyte expansion in culture. Effect of TGF beta-2, bFGF and L-ascorbic acid on bovine articular chondrocytes.. Acta Orthop Scand 1999 Feb;70(1):55-61.
- Pei M, Seidel J, Vunjak-Novakovic G, Freed LE. Growth factors for sequential cellular de- and re-differentiation in tissue engineering.. Biochem Biophys Res Commun 2002 May 31;294(1):149-54.
- Barbero A, Grogan S, Schäfer D, Heberer M, Mainil-Varlet P, Martin I. Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity.. Osteoarthritis Cartilage 2004 Jun;12(6):476-84.
- Chandrasekhar S, Harvey AK, Hrubey PS, Bendele AM. Arthritis induced by interleukin-1 is dependent on the site and frequency of intraarticular injection.. Clin Immunol Immunopathol 1990 Jun;55(3):382-400.
- Chubinskaya S, Huch K, Mikecz K, Cs-Szabo G, Hasty KA, Kuettner KE, Cole AA. Chondrocyte matrix metalloproteinase-8: up-regulation of neutrophil collagenase by interleukin-1 beta in human cartilage from knee and ankle joints.. Lab Invest 1996 Jan;74(1):232-40.
- Cui W, Akhurst RJ. Transforming Growth Factors ßs: Biochemistry and Biological Activities In-Vitro and In-Vivo. Growth Factors and Cytokines in Health and Disease Volume 1 B. Greenwich,USA, JAI Press Inc.; 1996. pp. 319–356.
- Vivien D, Redini F, Galéra P, Lebrun E, Loyau G, Pujol JP. Rabbit articular chondrocytes (RAC) express distinct transforming growth factor-beta receptor phenotypes as a function of cell cycle phases.. Exp Cell Res 1993 Mar;205(1):165-70.
- Fortier LA, Nixon AJ, Mohammed HO, Lust G. Altered biological activity of equine chondrocytes cultured in a three-dimensional fibrin matrix and supplemented with transforming growth factor beta-1.. Am J Vet Res 1997 Jan;58(1):66-70.
- Glansbeek HL, van der Kraan PM, Vitters EL, van den Berg WB. Correlation of the size of type II transforming growth factor beta (TGF-beta) receptor with TGF-beta responses of isolated bovine articular chondrocytes.. Ann Rheum Dis 1993 Nov;52(11):812-6.
- Jakob M, Démarteau O, Schäfer D, Hintermann B, Dick W, Heberer M, Martin I. Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro.. J Cell Biochem 2001 Mar 26;81(2):368-77.
- Desjardins MR, Hurtig MB. Diagnosis of equine stifle joint disorders: three cases.. Can Vet J 1991 Sep;32(9):543-50.
- Fortier LA, Mohammed HO, Lust G, Nixon AJ. Insulin-like growth factor-I enhances cell-based repair of articular cartilage.. J Bone Joint Surg Br 2002 Mar;84(2):276-88.
- Hurtig MB, Novak K, McPherson R, McFadden S, McGann LE, Mul drew K, Schachar NS. Osteochondral dowel transplantation for repair of focal defects in the knee: an outcome study using an ovine model.. Vet Surg 1998 Jan-Feb;27(1):5-16.
- Diaz-Romero J, Gaillard JP, Grogan SP, Nesic D, Trub T, Mainil-Varlet P. Immunophenotypic analysis of human articular chondrocytes: changes in surface markers associated with cell expansion in monolayer culture.. J Cell Physiol 2005 Mar;202(3):731-42.
- Eger W, Schumacher BL, Mollenhauer J, Kuettner KE, Cole AA. Human knee and ankle cartilage explants: catabolic differences.. J Orthop Res 2002 May;20(3):526-34.
- Elisseeff J, McIntosh W, Anseth K, Riley S, Ragan P, Langer R. Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks.. J Biomed Mater Res 2000 Aug;51(2):164-71.
- Bacenková D, Rosocha J, Svihla R, Vasko G, Bodnár J. [Repair of chondral defects of the knee using a combination of autologous chondrocytes and osteochondral allograft--an animal model. Part I: in vitro culture of autologous chondrocytes].. Acta Chir Orthop Traumatol Cech 2001;68(6):363-8.
- Nixon AJ, Lust G, Vernier-Singer M. Isolation, propagation, and cryopreservation of equine articular chondrocytes.. Am J Vet Res 1992 Dec;53(12):2364-70.
- Gravert HO. Rinderzucht. Einführung in die Züchtung, Fütterung und Haltung landwirtschaftlicher Nutztiere Hamburg und Berlin, Paul Parey; 1979. pp. 121–125.
- Getty R. The Anatomy of the Domestic Animals. 5th. Vol. 1. Philadelphia, W.B. Saunders Company; 1975.
- Habermehl KH. Die Altersbestimmung bei Haus- und Labortieren. 2nd. Berlin und Hamburg, Paul Parey; 1975. p. 214.
- Osborn KD, Trippel SB, Mankin HJ. Growth factor stimulation of adult articular cartilage.. J Orthop Res 1989;7(1):35-42.
- Scheerlinck JP. Functional and structural comparison of cytokines in different species.. Vet Immunol Immunopathol 1999 Dec 15;72(1-2):39-44.
- Glansbeek HL, van der Kraan PM, Vitters EL, van den Berg WB. Variable TGF-beta receptor expression regulates TGF-beta responses of articular chondrocytes.. Agents Actions Suppl 1993;39:139-45.
- Okazaki R, Sakai A, Nakamura T, Kunugita N, Norimura T, Suzuki K. Effects of transforming growth factor beta s and basic fibroblast growth factor on articular chondrocytes obtained from immobilised rabbit knees.. Ann Rheum Dis 1996 Mar;55(3):181-6.
- Zhang W, Liu HT. MAPK signal pathways in the regulation of cell proliferation in mammalian cells.. Cell Res 2002 Mar;12(1):9-18.
Citations
This article has been cited 7 times.- Storch C, Fuhrmann H, Schoeniger A. HOX Gene Expressions in Cultured Articular and Nasal Equine Chondrocytes. Animals (Basel) 2021 Aug 30;11(9).
- Cheng NC, Estes BT, Young TH, Guilak F. Engineered cartilage using primary chondrocytes cultured in a porous cartilage-derived matrix. Regen Med 2011 Jan;6(1):81-93.
- Mancuso L, Liuzzo MI, Fadda S, Pisu M, Cincotti A, Arras M, La Nasa G, Concas A, Cao G. In vitro ovine articular chondrocyte proliferation: experiments and modelling. Cell Prolif 2010 Jun;43(3):310-20.
- Ramaswamy S, Greco JB, Uluer MC, Zhang Z, Zhang Z, Fishbein KW, Spencer RG. Magnetic resonance imaging of chondrocytes labeled with superparamagnetic iron oxide nanoparticles in tissue-engineered cartilage. Tissue Eng Part A 2009 Dec;15(12):3899-910.
- Ng KW, Ateshian GA, Hung CT. Zonal chondrocytes seeded in a layered agarose hydrogel create engineered cartilage with depth-dependent cellular and mechanical inhomogeneity. Tissue Eng Part A 2009 Sep;15(9):2315-24.
- Schulze-Tanzil G, Müller RD, Kohl B, Schneider N, Ertel W, Ipaktchi K, Hünigen H, Gemeinhardt O, Stark R, John T. Differing in vitro biology of equine, ovine, porcine and human articular chondrocytes derived from the knee joint: an immunomorphological study. Histochem Cell Biol 2009 Feb;131(2):219-29.
- Porter A, Wang L, Han L, Lu XL. Bio-orthogonal Click Chemistry Methods to Evaluate the Metabolism of Inflammatory Challenged Cartilage after Traumatic Overloading. ACS Biomater Sci Eng 2022 Jun 13;8(6):2564-2573.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists