CD14-negative isolation enhances chondrogenesis in synovial fibroblasts.
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Journal: 2010/January - Tissue Engineering - Part A.
ISSN: 1937-335X
Abstract:
Synovial membrane has been shown to contain mesenchymal stem cells. We hypothesized that an enriched population of synovial fibroblasts would undergo chondrogenic differentiation and secrete cartilage extracellular matrix to a greater extent than would a mixed synovial cell population (MSCP). The optimum doses of transforming growth factor beta 1 (TGF-beta1) and insulin-like growth factor 1 (IGF-1) for chondrogenesis were investigated. CD14-negative isolation was used to obtain a porcine cell population enriched in type-B synovial fibroblasts (SFB) from an MSCP. The positive cell surface markers in SFB were CD90, CD44, and cadherin-11. SFB and MSCP were cultured in the presence of 20 ng/mL TGF-beta1 for 7 days, and SFB were demonstrated to have higher chondrogenic potential. Further dose-response studies were carried out using the SFB cells and several doses of TGF-beta1 (2, 10, 20, and 40 ng/mL) and/or IGF-1 (1, 10, 100, and 500 ng/mL) for 14 days. TGF-beta1 supplementation was essential for chondrogenesis and prevention of cell death, whereas IGF-1 did not have a significant effect on the SFB cell number or glycosaminoglycan production. This study demonstrates that the CD14-negative isolation yields an enhanced cell population SFB that is more potent than MSCP as a cell source for cartilage tissue engineering.
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Tissue Eng Part A 15(11): 3261-3270
PMID: 19382853

CD14-Negative Isolation Enhances Chondrogenesis in Synovial Fibroblasts

Department of Orthopaedics, Alpert Medical School of Brown University and Center for Restorative and Regenerative Medicine, Providence VA Medical Center and Brown University, Providence, Rhode Island.
Department of Orthopaedics, Alpert Medical School of Brown University, Providence, Rhode Island.
Department of Orthopaedics, West Virginia University, Morgantown, West Virginia.
Corresponding author.
Address correspondence to: Deborah McK. Ciombor, Ph.D., Department of Orthopaedics, Alpert Medical School of Brown University and Center for Restorative and Regenerative Medicine, Providence VA Medical Center and Brown University, Coro West Suite 402 C, 1 Hoppin St., Providence, RI 02903. E-mail:ude.nworb@robmoic_harobed
Address correspondence to: Deborah McK. Ciombor, Ph.D., Department of Orthopaedics, Alpert Medical School of Brown University and Center for Restorative and Regenerative Medicine, Providence VA Medical Center and Brown University, Coro West Suite 402 C, 1 Hoppin St., Providence, RI 02903. E-mail:ude.nworb@robmoic_harobed
Received 2008 May 8; Accepted 2009 Apr 21.
Copyright 2009, Mary Ann Liebert, Inc.

Abstract

Synovial membrane has been shown to contain mesenchymal stem cells. We hypothesized that an enriched population of synovial fibroblasts would undergo chondrogenic differentiation and secrete cartilage extracellular matrix to a greater extent than would a mixed synovial cell population (MSCP). The optimum doses of transforming growth factor beta 1 (TGF-β1) and insulin-like growth factor 1 (IGF-1) for chondrogenesis were investigated. CD14-negative isolation was used to obtain a porcine cell population enriched in type-B synovial fibroblasts (SFB) from an MSCP. The positive cell surface markers in SFB were CD90, CD44, and cadherin-11. SFB and MSCP were cultured in the presence of 20 ng/mL TGF-β1 for 7 days, and SFB were demonstrated to have higher chondrogenic potential. Further dose–response studies were carried out using the SFB cells and several doses of TGF-β1 (2, 10, 20, and 40 ng/mL) and/or IGF-1 (1, 10, 100, and 500 ng/mL) for 14 days. TGF-β1 supplementation was essential for chondrogenesis and prevention of cell death, whereas IGF-1 did not have a significant effect on the SFB cell number or glycosaminoglycan production. This study demonstrates that the CD14-negative isolation yields an enhanced cell population SFB that is more potent than MSCP as a cell source for cartilage tissue engineering.

Abstract

References

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