Influence of chondrocytes on the chondrogenic differentiation of adipose stem cells

Jai Sun Lee, Gun Il Im

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

In this study, whether or not chondrogenic differentiation of adipose-derived stem cells (ASCs) could be enhanced by soluble factors from or coculture with chondrocytes was determined. In vitro pellet cultures were carried out in five ways using ASCs or chondrocytes in passage 3 as follows: #1, 2.5×105 ASCs were cultured in Dulbecco's modified Eagle's medium/F-12 supplemented with 1% ITS, 10-7 M dexamethasone, 50μM ascorbate-2-phosphate, 50μM L-proline, and 1mM sodium pyruvate; #2, 2.5×105 chondrocytes were cultured in the same medium as #1: #3, 1.25×105 ASCs and 1.25×105 chondrocytes were mixed and cocultured in the same medium as #1; #4, 2.5×105 ASCs were cultured in a medium that was a 1:1 mixture of the same fresh medium as #1 and conditioned medium from chondrocyte culture (#2); #5, 2.5×105 ASCs were cultured in the same medium as #1 and 5ng/mL of transforming growth factor-β2 and 100ng/mL of BMP-7. After 3 weeks, the glycosaminoglycan level that normalized to the DNA amount was significantly increased by 25% in ASCs treated with condition medium from chondrocyte cultures (p=0.028) and by 37% in ASC-chondrocyte cocultures (p=0.042). The glycosaminoglycan level was 37% greater in chondrocytes (p=0.046) and 50% greater in ASCs cultured under growth factor cocktails than the control ASCs. The gene expression of SOX-9 significantly increased by >10-fold (p<0.05) in ASCs treated with the conditioned medium from chondrocyte cultures and ASC-chondrocyte cocultures compared with the control ASCs; whereas COL2A1 significantly increased ∼100-fold (p<0.05) in either condition. COL10A1 gene expression increased by treating either with conditioned medium or with coculture (p<0.05), but COL1A1 gene expression did not significantly change in either condition. Western blotting of SOX-9 and immunochemistry for types II, I, and X collagen largely parallel the results from gene expression studies. It is concluded that the signals from chondrocytes, in the form of soluble factors or by direct interaction, effectively promote chondrogenic differentiation of ASCs during in vitro pellet culture. This work may present a simple and innovative method for generating cartilaginous tissue from ASCs and shed a new light in cartilage tissue engineering from ASCs.

Original languageEnglish
Pages (from-to)3569-3577
Number of pages9
JournalTissue Engineering - Part A.
Volume16
Issue number12
DOIs
StatePublished - 1 Dec 2010

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