Adipose-derived mesenchymal stem cells from the sand rat: transforming growth factor beta and 3D co-culture with human disc cells stimulate proteoglycan and collagen type I rich extracellular matrix.
BACKGROUND
Adult mesenchymal stem cell therapy has a potential application in the biological treatment of disc degeneration. Our objectives were: to direct adipose-derived mesenchymal stem cells (AD-MSC) from the sand rat to produce a proteoglycan and collagen type I extracellular matrix (ECM) rich in known ECM components of the annulus fibrosis of disc; and to stimulate proteoglycan production by co-culture of human annulus cells with AD-MSC.


METHODS
AD-MSC were isolated and characterised by adherence to plastic, appropriate expression of cluster of differentiation (CD) markers, and differentiation to osteoblasts and chondrocytes in vitro. AD-MSC were grown in three-dimensional (3D) culture and treated with or without transforming growth factor beta (TGFbeta) to direct them to produce annulus-like ECM as determined by proteoglycan content and collagen expression. AD-MSC were co-cultured with human annulus cells and grown in 3D culture.


RESULTS
AD-MSC produced a proteoglycan and collagen type I rich ECM after treatment with TGFbeta in 3D culture as confirmed by a 48% increase in proteoglycan content assayed by 1,9-dimethylmethylene blue (DMB), and by immunohistochemical identification of ECM components. Co-culture of human annulus and sand rat AD-MSC in 3D culture resulted in a 20% increase in proteoglycan production compared with the predicted value of the sum of the individual cultures.


CONCLUSIONS
Results support the hypothesis that AD-MSC have potential in cell-based therapy for disc degeneration.
Tapp
Hazel
H
Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, NC 28232, USA. hazel.tapp@carolinashealthcare.org
Deepe
Ray
R
Ingram
Jane A
JA
Kuremsky
Marshall
M
Hanley
Edward N
EN
Gruber
Helen E
HE
D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002448:Cell Adhesion; D002454:Cell Differentiation; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D018920:Coculture Techniques; D024042:Collagen Type I / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005849:Gerbillinae; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats; D013122:Spinal Diseases / Q000628:therapy; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D024042:Collagen Type I; D011509:Proteoglycans; D016212:Transforming Growth Factor beta