A Comparison of Three-dimensional Culture Systems to Evaluate in Vitro Chondrogenesis of Equine Bone Marrow-derived Mesenchymal Stem Cells

Overview

Journal Tissue Eng Part A

Specialties Anatomy & Histology
Biomedical Engineering
Biotechnology

Date 2013 Jun 4

PMID 23725547

Citations 29

Authors

Ashlee E Watts

Jeremy C Ackerman-Yost

Alan J Nixon

Affiliations

Soon will be listed here.

Abstract

Objective: To compare in vitro three-dimensional (3D) culture systems that model chondrogenesis of bone marrow-derived mesenchymal stem cells (MSCs).

Methods: MSCs from five horses 2-3 years of age were consolidated in fibrin 0.3% alginate, 1.2% alginate, 2.5×10(5) cell pellets, 5×10(5) cell pellets, and 2% agarose, and maintained in chondrogenic medium with supplemental TGF-β1 for 4 weeks. Pellets and media were tested at days 1, 14, and 28 for gene expression of markers of chondrogenic maturation and hypertrophy (ACAN, COL2B, COL10, SOX9, 18S), and evaluated by histology (hematoxylin and eosin, Toluidine Blue) and immunohistochemistry (collagen type II and X).

Results: alginate, fibrin alginate (FA), and both pellet culture systems resulted in chondrogenic transformation. Adequate RNA was not obtained from agarose cultures at any time point. There was increased COL2B, ACAN, and SOX9 expression on day 14 from both pellet culture systems. On day 28, increased expression of COL2B was maintained in 5×10(5) cell pellets and there was no difference in ACAN and SOX9 between FA and both pellet cultures. COL10 expression was significantly lower in FA cultures on day 28. Collagen type II was abundantly formed in all culture systems except alginate and collagen type X was least in FA hydrogels.

Conclusion: equine MSCs respond to 3D culture in FA blended hydrogel and both pellet culture systems with chondrogenic induction. For prevention of terminal differentiation and hypertrophy, FA culture may be superior to pellet culture systems.

Citing Articles

The systemic cellular immune response against allogeneic mesenchymal stem cells is influenced by inflammation, differentiation and MHC compatibility: study in the horse.

Cequier A, Vazquez F, Vitoria A, Bernad E, Fuente S, Serrano M Front Vet Sci. 2024; 11:1391872.

PMID: 38957800 PMC: 11217187. DOI: 10.3389/fvets.2024.1391872.

NOS inhibition reverses TLR2-induced chondrocyte dysfunction and attenuates age-related osteoarthritis.

Shen P, Serve S, Wu P, Liu X, Dai Y, Duran-Hernandez N Proc Natl Acad Sci U S A. 2023; 120(29):e2207993120.

PMID: 37428931 PMC: 10629581. DOI: 10.1073/pnas.2207993120.

Human 3D nucleus pulposus microtissue model to evaluate the potential of pre-conditioned nasal chondrocytes for the repair of degenerated intervertebral disc.

Kasamkattil J, Gryadunova A, Schmid R, Gay-Dujak M, Dasen B, Hilpert M Front Bioeng Biotechnol. 2023; 11:1119009.

PMID: 36865027 PMC: 9971624. DOI: 10.3389/fbioe.2023.1119009.

Controlling Microenvironments with Organs-on-Chips for Osteoarthritis Modelling.

Ong L, Fan X, Sun A, Mei L, Toh Y, Prasadam I Cells. 2023; 12(4).

PMID: 36831245 PMC: 9954502. DOI: 10.3390/cells12040579.

Performance of Colombian Silk Fibroin Hydrogels for Hyaline Cartilage Tissue Engineering.

Zuluaga-Velez A, Toro-Acevedo C, Quintero-Martinez A, Melchor-Moncada J, Pedraza-Ordonez F, Aguilar-Fernandez E J Funct Biomater. 2022; 13(4).

PMID: 36547557 PMC: 9788426. DOI: 10.3390/jfb13040297.

References

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; 331(14):889-95. DOI: 10.1056/NEJM199410063311401. View

Dresdale A, Rose E, Jeevanandam V, REEMTSMA K, Bowman F, Malm J . Preparation of fibrin glue from single-donor fresh-frozen plasma. Surgery. 1985; 97(6):750-5. View

Hyc A, Malejczyk J, Osiecka A, Moskalewski S . Immunological response against allogeneic chondrocytes transplanted into joint surface defects in rats. Cell Transplant. 1997; 6(2):119-24. DOI: 10.1177/096368979700600205. View

Hendrickson D, Nixon A, Grande D, Todhunter R, Minor R, Erb H . Chondrocyte-fibrin matrix transplants for resurfacing extensive articular cartilage defects. J Orthop Res. 1994; 12(4):485-97. DOI: 10.1002/jor.1100120405. View

Hunter D . Pharmacologic therapy for osteoarthritis--the era of disease modification. Nat Rev Rheumatol. 2010; 7(1):13-22. DOI: 10.1038/nrrheum.2010.178. View

Jeffcott L, Rossdale P, Freestone J, Frank C . An assessment of wastage in thoroughbred racing from conception to 4 years of age. Equine Vet J. 1982; 14(3):185-98. DOI: 10.1111/j.2042-3306.1982.tb02389.x. View

Matsumoto T, Okabe T, Ikawa T, Iida T, Yasuda H, Nakamura H . Articular cartilage repair with autologous bone marrow mesenchymal cells. J Cell Physiol. 2010; 225(2):291-5. DOI: 10.1002/jcp.22223. View

Lefebvre V, Smits P . Transcriptional control of chondrocyte fate and differentiation. Birth Defects Res C Embryo Today. 2005; 75(3):200-12. DOI: 10.1002/bdrc.20048. View

Wilke M, Nydam D, Nixon A . Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model. J Orthop Res. 2007; 25(7):913-25. DOI: 10.1002/jor.20382. View

10.

Sams A, Nixon A . Chondrocyte-laden collagen scaffolds for resurfacing extensive articular cartilage defects. Osteoarthritis Cartilage. 1995; 3(1):47-59. DOI: 10.1016/s1063-4584(05)80037-8. View

11.

Fortier L, Potter H, Rickey E, Schnabel L, Foo L, Chong L . Concentrated bone marrow aspirate improves full-thickness cartilage repair compared with microfracture in the equine model. J Bone Joint Surg Am. 2010; 92(10):1927-37. DOI: 10.2106/JBJS.I.01284. View

12.

Kasashima Y, Ueno T, Tomita A, Goodship A, Smith R . Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells. Equine Vet J. 2011; 43(3):288-94. DOI: 10.1111/j.2042-3306.2010.00215.x. View

13.

Elves M . A study of the transplantation antigens on chondrocytes from articular cartilage. J Bone Joint Surg Br. 1974; 56(1):178-85. View

14.

Strauss E, Goodrich L, Chen C, Hidaka C, Nixon A . Biochemical and biomechanical properties of lesion and adjacent articular cartilage after chondral defect repair in an equine model. Am J Sports Med. 2005; 33(11):1647-53. DOI: 10.1177/0363546505275487. View

15.

Tuan R . Biology of developmental and regenerative skeletogenesis. Clin Orthop Relat Res. 2004; (427 Suppl):S105-17. DOI: 10.1097/01.blo.0000143560.41767.ee. View

16.

Yang I, Kim S, Kim Y, Sun H, Kim S, Lee J . Comparison of phenotypic characterization between "alginate bead" and "pellet" culture systems as chondrogenic differentiation models for human mesenchymal stem cells. Yonsei Med J. 2004; 45(5):891-900. DOI: 10.3349/ymj.2004.45.5.891. View

17.

Johnstone B, Hering T, Caplan A, Goldberg V, Yoo J . In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res. 1998; 238(1):265-72. DOI: 10.1006/excr.1997.3858. View

18.

McIlwraith C, Frisbie D, Rodkey W, Kisiday J, Werpy N, Kawcak C . Evaluation of intra-articular mesenchymal stem cells to augment healing of microfractured chondral defects. Arthroscopy. 2011; 27(11):1552-61. DOI: 10.1016/j.arthro.2011.06.002. View

19.

Ortved K, Nixon A, Mohammed H, Fortier L . Treatment of subchondral cystic lesions of the medial femoral condyle of mature horses with growth factor enhanced chondrocyte grafts: a retrospective study of 49 cases. Equine Vet J. 2011; 44(5):606-13. DOI: 10.1111/j.2042-3306.2011.00510.x. View

20.

Ichinose S, Tagami M, Muneta T, Sekiya I . Morphological examination during in vitro cartilage formation by human mesenchymal stem cells. Cell Tissue Res. 2005; 322(2):217-26. DOI: 10.1007/s00441-005-1140-6. View