Skip to main content
Top
Published in: International Orthopaedics 4/2013

Open Access 01-04-2013 | Original Paper

Co-cultivated mesenchymal stem cells support chondrocytic differentiation of articular chondrocytes

Authors: Qiang Zuo, Weiding Cui, Feng Liu, Qing Wang, Zhefeng Chen, Weimin Fan

Published in: International Orthopaedics | Issue 4/2013

Login to get access

Abstract

Purpose

This study investigated which of the reciprocal stimuli between articular chondrocytes (ACs) and mesenchymal stem cells (MSCs) played the more important role in enhancing cartilage matrix formation, and examined the relative importance of physical contact and soluble factors in the co-culture system.

Methods

Rat ACs and bone marrow MSCs with green fluorescent protein (GFP-BMSCs) were co-cultured in vitro with or without direct cell–cell contact at the ratio of 2:1. After co-culturing in direct cell–cell contact, ACs and GFP-BMSCs were separated by flow cytometry. The effects of different co-culture methods were analysed by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and western blotting.

Results

SOX-9, COL2 and aggrecan mRNA levels and protein expression in ACs co-cultured with direct cell–cell contact were significantly higher than in ACs co-cultured without direct cell–cell contact; and similar results were found in GFP-BMSCs. After co-culture either with or without direct cell–cell contact, mRNA levels and protein expression of SOX-9, COL2 and aggrecan in GFP-BMSCs were significantly lower than in ACs in the equivalent co-culture systems. Though the expression of chondrocyte-specific proteins in GFP-BMSCs was enhanced, the protein expression was still much lower than in ACs cultured alone.

Conclusions

Reciprocal interactions exist between ACs and BMSCs in co-culture. The stimulating and supporting effects of BMSCs on ACs were more important in enhancing cartilage-matrix formation than the reciprocal effect of ACs on BMSCs. Both soluble factors and direct physical contact occur in AC/BMSC co-cultures, with physical contact playing a predominant, or at least very important role.
Literature
1.
go back to reference Mo XT, Guo SC, Xie HQ et al (2009) Variations in the ratios of co-cultured mesenchymal stem cells and chondrocytes regulate the expression of cartilaginous and osseous phenotype in alginate constructs. Bone 45(1):42–51PubMedCrossRef Mo XT, Guo SC, Xie HQ et al (2009) Variations in the ratios of co-cultured mesenchymal stem cells and chondrocytes regulate the expression of cartilaginous and osseous phenotype in alginate constructs. Bone 45(1):42–51PubMedCrossRef
2.
go back to reference Gruber HE, Deepe R, Hoelscher GL et al (2010) Human adipose-derived mesenchymal stem cells: direction to a phenotype sharing similarities with the disc, gene expression profiling, and coculture with human annulus cells. Tissue Eng Part A 16(9):2843–2860PubMedCrossRef Gruber HE, Deepe R, Hoelscher GL et al (2010) Human adipose-derived mesenchymal stem cells: direction to a phenotype sharing similarities with the disc, gene expression profiling, and coculture with human annulus cells. Tissue Eng Part A 16(9):2843–2860PubMedCrossRef
3.
go back to reference Chen WH, Lai MT, Wu AT et al (2009) In vitro stage-specific chondrogenesis of mesenchymal stem cells committed to chondrocytes. Arthritis Rheum 60(2):450–459PubMedCrossRef Chen WH, Lai MT, Wu AT et al (2009) In vitro stage-specific chondrogenesis of mesenchymal stem cells committed to chondrocytes. Arthritis Rheum 60(2):450–459PubMedCrossRef
4.
go back to reference Bigdeli N, Karlsson C, Strehl R et al (2009) Coculture of human embryonic stem cells and human articular chondrocytes results in significantly altered phenotype and improved chondrogenic differentiation. Stem Cells 27(8):1812–1821PubMedCrossRef Bigdeli N, Karlsson C, Strehl R et al (2009) Coculture of human embryonic stem cells and human articular chondrocytes results in significantly altered phenotype and improved chondrogenic differentiation. Stem Cells 27(8):1812–1821PubMedCrossRef
5.
go back to reference Richardson SM, Walker RV, Parker S et al (2006) Intervertebral disc cell-mediated mesenchymal stem cell differentiation. Stem Cells 24(3):707–716PubMedCrossRef Richardson SM, Walker RV, Parker S et al (2006) Intervertebral disc cell-mediated mesenchymal stem cell differentiation. Stem Cells 24(3):707–716PubMedCrossRef
6.
go back to reference Yang YH, Lee AJ, Barabino GA (2012) Coculture-driven mesenchymal stem cell-differentiated articular chondrocyte-like cells support neocartilage development. Stem Cells Transl Med 1(11):843–854PubMedCrossRef Yang YH, Lee AJ, Barabino GA (2012) Coculture-driven mesenchymal stem cell-differentiated articular chondrocyte-like cells support neocartilage development. Stem Cells Transl Med 1(11):843–854PubMedCrossRef
7.
go back to reference Kim JS, Ryoo ZY, Chun JS (2007) Cytokine-like 1 (Cytl1) regulates the chondrogenesis of mesenchymal cells. J Biol Chem 282(40):29359–29367PubMedCrossRef Kim JS, Ryoo ZY, Chun JS (2007) Cytokine-like 1 (Cytl1) regulates the chondrogenesis of mesenchymal cells. J Biol Chem 282(40):29359–29367PubMedCrossRef
8.
go back to reference Fischer J, Dickhut A, Rickert M et al (2010) Human articular chondrocytes secrete parathyroid hormone-related protein and inhibit hypertrophy of mesenchymal stem cells in coculture during chondrogenesis. Arthritis Rheum 62(9):2696–2706PubMedCrossRef Fischer J, Dickhut A, Rickert M et al (2010) Human articular chondrocytes secrete parathyroid hormone-related protein and inhibit hypertrophy of mesenchymal stem cells in coculture during chondrogenesis. Arthritis Rheum 62(9):2696–2706PubMedCrossRef
9.
go back to reference Keller B, Yang T, Chen Y et al (2011) Interaction of TGFbeta and BMP signaling pathways during chondrogenesis. PLoS One 6(1):e16421PubMedCrossRef Keller B, Yang T, Chen Y et al (2011) Interaction of TGFbeta and BMP signaling pathways during chondrogenesis. PLoS One 6(1):e16421PubMedCrossRef
10.
go back to reference Wu L, Leijten JC, Georgi N et al (2011) Trophic effects of mesenchymal stem cells increase chondrocyte proliferation and matrix formation. Tissue Eng Part A 17(9–10):1425–1436PubMedCrossRef Wu L, Leijten JC, Georgi N et al (2011) Trophic effects of mesenchymal stem cells increase chondrocyte proliferation and matrix formation. Tissue Eng Part A 17(9–10):1425–1436PubMedCrossRef
11.
go back to reference Hwang NS, Varghese S, Puleo C et al (2007) Morphogenetic signals from chondrocytes promote chondrogenic and osteogenic differentiation of mesenchymal stem cells. J Cell Physiol 212(2):281–284PubMedCrossRef Hwang NS, Varghese S, Puleo C et al (2007) Morphogenetic signals from chondrocytes promote chondrogenic and osteogenic differentiation of mesenchymal stem cells. J Cell Physiol 212(2):281–284PubMedCrossRef
12.
go back to reference Hwang NS, Im SG, Wu PB et al (2011) Chondrogenic priming adipose-mesenchymal stem cells for cartilage tissue regeneration. Pharm Res 28(6):1395–1405PubMedCrossRef Hwang NS, Im SG, Wu PB et al (2011) Chondrogenic priming adipose-mesenchymal stem cells for cartilage tissue regeneration. Pharm Res 28(6):1395–1405PubMedCrossRef
13.
go back to reference Aung A, Gupta G, Majid G et al (2011) Osteoarthritic chondrocyte-secreted morphogens induce chondrogenic differentiation of human mesenchymal stem cells. Arthritis Rheum 63(1):148–158PubMedCrossRef Aung A, Gupta G, Majid G et al (2011) Osteoarthritic chondrocyte-secreted morphogens induce chondrogenic differentiation of human mesenchymal stem cells. Arthritis Rheum 63(1):148–158PubMedCrossRef
14.
go back to reference Yamamoto Y, Mochida J, Sakai D et al (2004) Upregulation of the viability of nucleus pulposus cells by bone marrow-derived stromal cells: significance of direct cell-to-cell contact in coculture system. Spine (Phila Pa 1976) 29(14):1508–1514CrossRef Yamamoto Y, Mochida J, Sakai D et al (2004) Upregulation of the viability of nucleus pulposus cells by bone marrow-derived stromal cells: significance of direct cell-to-cell contact in coculture system. Spine (Phila Pa 1976) 29(14):1508–1514CrossRef
15.
go back to reference Heng BC, Cao T, Lee EH (2004) Directing stem cell differentiation into the chondrogenic lineage in vitro. Stem Cells 22(7):1152–1167PubMedCrossRef Heng BC, Cao T, Lee EH (2004) Directing stem cell differentiation into the chondrogenic lineage in vitro. Stem Cells 22(7):1152–1167PubMedCrossRef
16.
go back to reference Qing C, Wei-ding C, Wei-min F (2011) Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components. Braz J Med Biol Res 44(4):303–310PubMed Qing C, Wei-ding C, Wei-min F (2011) Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components. Braz J Med Biol Res 44(4):303–310PubMed
17.
go back to reference Wei A, Chung SA, Tao H et al (2009) Differentiation of rodent bone marrow mesenchymal stem cells into intervertebral disc-like cells following coculture with rat disc tissue. Tissue Eng Part A 15(9):2581–2595PubMedCrossRef Wei A, Chung SA, Tao H et al (2009) Differentiation of rodent bone marrow mesenchymal stem cells into intervertebral disc-like cells following coculture with rat disc tissue. Tissue Eng Part A 15(9):2581–2595PubMedCrossRef
18.
go back to reference Schilling T, Noth U, Klein-Hitpass L et al (2007) Plasticity in adipogenesis and osteogenesis of human mesenchymal stem cells. Mol Cell Endocrinol 271(1–2):1–17PubMedCrossRef Schilling T, Noth U, Klein-Hitpass L et al (2007) Plasticity in adipogenesis and osteogenesis of human mesenchymal stem cells. Mol Cell Endocrinol 271(1–2):1–17PubMedCrossRef
19.
go back to reference Johnstone B, Hering TM, Caplan AI et al (1998) In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 238(1):265–272PubMedCrossRef Johnstone B, Hering TM, Caplan AI et al (1998) In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 238(1):265–272PubMedCrossRef
20.
go back to reference Liu X, Sun H, Yan D et al (2010) In vivo ectopic chondrogenesis of BMSCs directed by mature chondrocytes. Biomaterials 31(36):9406–9414PubMedCrossRef Liu X, Sun H, Yan D et al (2010) In vivo ectopic chondrogenesis of BMSCs directed by mature chondrocytes. Biomaterials 31(36):9406–9414PubMedCrossRef
Metadata
Title
Co-cultivated mesenchymal stem cells support chondrocytic differentiation of articular chondrocytes
Authors
Qiang Zuo
Weiding Cui
Feng Liu
Qing Wang
Zhefeng Chen
Weimin Fan
Publication date
01-04-2013
Publisher
Springer-Verlag
Published in
International Orthopaedics / Issue 4/2013
Print ISSN: 0341-2695
Electronic ISSN: 1432-5195
DOI
https://doi.org/10.1007/s00264-013-1782-z

Other articles of this Issue 4/2013

International Orthopaedics 4/2013 Go to the issue