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In vitro interaction between muscle-derived stem cells and nucleus pulposus cells

Vadalà, G and Sobajima, S and Lee, JY and Huard, J and Denaro, V and Kang, JD and Gilbertson, LG (2008) In vitro interaction between muscle-derived stem cells and nucleus pulposus cells. Spine Journal, 8 (5). 804 - 809. ISSN 1529-9430

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Abstract

Background context: Current therapies for intervertebral disc degeneration (IDD) are aimed at treating the clinical symptoms arising from IDD rather than directly treating the underlying problem. Pathophysiology of IDD is characterized by a progressive decrease in proteoglycan content and cell density in the nucleus pulposus (NP). A cell-based therapy is a promising concept that uses various cell types to repopulate the disc in an attempt to slow, stop, or reverse the progressive loss of proteoglycans. Stem cells appear to be an excellent candidate for this purpose, based on their ability to differentiate into various connective tissue lineages. The muscle tissue could serve as a good source of adult stem cells because of its vast abundance through out the human body. Purpose: To examine the interaction between the nucleus pulposus cells (NPCs) and the muscle-derived stem cells (MDSCs) in vitro. Study design: NPCs and MDSCs were cocultured and proteoglycan production and cell proliferation were evaluated. Methods: Various ratios of human NPCs were cocultured for 2 weeks with murine MDSCs (transduced with retro/LacZ) in a monolayer culture. Each well contained an admixture of cells with NPC-to-MDSC ratios of 0:100, 25:75, 50:50, 75:25, 100:0. Glycosaminoglycan (GAG) content (1,9 dimethylmethylene blue [DMMB]), newly synthesized proteoglycan (35S incorporation), and DNA content were measured, and cultures were stained with 5-bromo-4-chloro-3-indolyl-β-D-galactosidase (X-Gal) for cell counting. Results: The NPC-to-MDSC ratio of 75:25 resulted in a significant increase in GAG content compared with NPCs alone. All coculture ratios showed increase in GAG content in comparison with MDSC culture alone. In addition, cocultures showed a significant increase in 35S incorporation normalized to DNA content in comparison with MDSC alone. Conclusions: The data from this study shows a synergistic effect between MDSCs and NPCs resulting in an upregulated proteoglycan synthesis and NPCs proliferation in vitro. © 2008 Elsevier Inc. All rights reserved.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Vadalà, G
Sobajima, S
Lee, JY
Huard, J
Denaro, V
Kang, JDjdkang@pitt.eduJDKANG
Gilbertson, LG
Centers: Other Centers, Institutes, Offices, or Units > Stem Cell Research Center
Date: 1 September 2008
Date Type: Publication
Journal or Publication Title: Spine Journal
Volume: 8
Number: 5
Page Range: 804 - 809
DOI or Unique Handle: 10.1016/j.spinee.2007.07.394
Schools and Programs: School of Medicine > Orthopaedic Surgery
Refereed: Yes
ISSN: 1529-9430
MeSH Headings: Adult Stem Cells--cytology; Adult Stem Cells--metabolism; Animals; Cell Differentiation--physiology; Cell Proliferation; Coculture Techniques; Extracellular Matrix--chemistry; Extracellular Matrix--metabolism; Extracellular Matrix Proteins--metabolism; Female; Glycosaminoglycans--biosynthesis; Humans; Intervertebral Disc--cytology; Intervertebral Disc--metabolism; Intervertebral Disc Displacement--therapy; Mice; Muscle, Skeletal--cytology; Proteoglycans--biosynthesis; Tissue Engineering--methods; Transduction, Genetic
PubMed ID: 18023623
Date Deposited: 15 May 2014 20:26
Last Modified: 02 Feb 2019 21:55
URI: http://d-scholarship.pitt.edu/id/eprint/21600

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