Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement

Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.


Study of Chondrogenic Potential of Subpopulations of Cells Expressing Mesenchymal Stem Cell Markers Derived from Human Synovial Membranes.

Arufe4,  María C., De la Fuente3,  Alexandre, Diaz2,  Silvia, Fuentes2,  Isaac, de Toro1,  Francisco J., Blanco5,  Francisco J.

Dpto. Medicine, Area of Anatomy and Human Embryology, Campus Oza s/n. Fac. of Health Science, University of A Coruna, CIBER-BBN, Instituto de Salud Carlos III, Spain
Osteoarticular and Aging Research Lab. Cellular Therapy Unit, INIBIC-CHUAC, A Coruña, Dpto. Medicine, Area of Anatomy and Human Embryology, Campus Oza s/n. Fac. of Health Science, University of A Coruna, CIBER-BBN, Instituto
Osteoarticular and Aging Research Lab. Cellular Therapy Unit, INIBIC-CHUAC, A Coruña, Spain, CIBER-BBN, Instituto de Salud Carlos III, Spain
Osteoarticular and Aging Research Lab. Cellular Therapy Unit. INIBIC-CHUAC, A Coruna, Dpto. Medicine, Area of Anatomy and Human Embryology, Campus Oza s/n. Fac. of Health Science, University of A Coruna, CIBER-BBN, Instituto de Sal
Osteoarticular and Aging Research Lab. Cellular Therapy Unit, INIBIC-CHUAC, A Coruña.Cathedra BIOIBERICA of Cell Therapy University of A Coruna, A Coruña, Spain, CIBER-BBN, Instituto de Salud Carlos III, Spain

Introduction:

Synovial membrane mesenchymal stem cells (MSCs) have been demonstrated to be a good source of cells for the study of cartilage tissue engineering. Multiple stem cells markers have been found by flow cytometry and immunofluorescence in MSCs from human synovial membrane pools. In this study we analyzed the chondrogenic potential of subpopulations of MSCs derived from human synovial membranes enriched for CD73, CD106 and CD271 markers.

Material and Methods:

Subpopulations of human synovial membrane MSCs enriched for CD73, CD106 and CD271 markers were isolated using a cytometry sorter and characterized by flow cytometry for MSC markers. The expression of Sox9, Nanog and Runx2 genes by these cells was measured by reverse transcriptase-polymerase chain reaction. The chondrogenesis of each subpopulation was assessed by culturing the cells in a defined medium to produce spontaneous spheroid formation and differentiation towards chondrocyte-like cells. The examination of the spheroids by histological and immunohistochemical analyses for collagen type II (COL2), aggrecan, collagen type I (COL1), metalloprotease 13 (MMP13) and collagen type × (COLX) levels were performed to assess their chondrogenesis capacity. The adipogenesis and osteogenesis potential of each subpopulation was determined using commercial media; the resulting cells were stained with oil red O or red alizarin to test the degree of differentiation.

Results:

The subpopulations had different profiles of cells positive for the MSC markers CD44, CD69, CD73, CD90 and CD105 and showed different expression levels of the genes Sox9, Nanog, Runx2 involved in chondrogenesis, undifferentiation and osteoblastogenesis, respectively. Immunohistochemical analysis demonstrated that COL1, COL2, COLX, MMP13 and aggrecan were expressed in the spheroids as soon as 14 days of culture. The CD271+ subpopulation expressed the highest levels of COL2 staining compared to the other subpopulations. CD105 and Runx2 were shown by immunohistochemistry and genetic analysis to have significantly higher expression CD271+ subpopulation than the other subpopulations.

Conclusions:

Spheroids formed from CD271-enriched and CD73-enriched MSCs from normal human synovial membranes mimic the native cartilage extracellular matrix more closely than CD106+ MSCs and are possible candidates for use in cartilage tissue engineering. Both cell types have potential for promoting the differentiation of MSCs into chondrocytes, presenting new possibilities for achieving intrinsic cartilage repair.

To cite this abstract, please use the following information:
Arufe, María C., De la Fuente, Alexandre, Diaz, Silvia, Fuentes, Isaac, de Toro, Francisco J., Blanco, Francisco J.; Study of Chondrogenic Potential of Subpopulations of Cells Expressing Mesenchymal Stem Cell Markers Derived from Human Synovial Membranes. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :1498
DOI: 10.1002/art.29264

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