Evaluation of adult equine bone marrow- and adipose-derived progenitor cell chondrogenesis in hydrogel cultures.
Bone marrow mesenchymal stem cells (BM-MSCs) and adipose-derived progenitor cells (ADPCs) are potential alternatives to autologous chondrocytes for cartilage resurfacing strategies. In this study, the chondrogenic potentials of these cell types were compared by quantifying neo-tissue synthesis and assaying gene expression and accumulation of extracellular matrix (ECM) components of cartilage. Adult equine progenitor cells encapsulated in agarose or self-assembling peptide hydrogels were cultured in the presence or absence of TGFbeta1 for 3 weeks. In BM-MSCs-seeded hydrogels, TGFbeta1 stimulated ECM synthesis and accumulation 3-41-fold relative to TGFbeta1-free culture. In ADPC cultures, TGFbeta1 stimulated a significant increase in ECM synthesis and accumulation in peptide (18-29-fold) but not agarose hydrogels. Chromatographic analysis of BM-MSC-seeded agarose and peptide hydrogels cultured in TGFbeta1 medium showed extensive synthesis of aggrecan-like proteoglycan monomers. ADPCs seeded in peptide hydrogel also synthesized aggrecan-like proteoglycans, although to a lesser extent than seen in BM-MSC hydrogels, whereas aggrecan-like proteoglycan synthesis in ADPC-seeded agarose was minimal. RT-PCR analysis of TGFbeta1 cultures showed detectable levels of type II collagen gene expression in BM-MSC but not ADPC cultures. Histological analysis of TGFbeta1-cultured peptide hydrogels showed the deposition of a continuous proteoglycan- and type II collagen rich ECM for BM-MSCs but not ADPCs. Therefore, this study showed both protein and gene expression evidence of superior chondrogenesis of BM-MSCs relative to ADPCs.
Kisiday
John D
JD
Orthopaedic Research Center, Department of Clinical Science, Colorado State University, 300 W. Drake Road, Fort Collins, Colorado 80523, USA. kisiday@lamar.colostate.edu
Kopesky
Paul W
PW
Evans
Christopher H
CH
Grodzinsky
Alan J
AJ
McIlwraith
C Wayne
CW
Frisbie
David D
DD
D000273:Adipose Tissue / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002449:Cell Aggregation; D002454:Cell Differentiation; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000502:physiology*; D002847:Chromatography, Agarose; D002850:Chromatography, Gel; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006736:Horses; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D012685:Sepharose; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
D053674:Aggrecans; D024042:Collagen Type I; D024043:Collagen Type II; D003470:Culture Media; D053773:Transforming Growth Factor beta1; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C043127:Superose 6b; D012685:Sepharose