Changes in chondrogenic phenotype and gene expression profiles associated with the in vitro expansion of human synovium‐derived cells

Abstract

We undertook this study to characterize changes in the proliferative capacities, chondrogenic phenotypes, and gene expression profiles of human synovium-derived progenitor cells from osteoarthritic patients during in vitro expansion. Cells isolated from osteoarthritic synovia were cultured, and growth rates during serial passages were evaluated. Surface molecule expressions were determined by flow cytometry and cytogenetic analyses were performed. After chondrogenic differentiation in cell pellets, we evaluated type II collagen and glycosaminoglycan (GAG) synthesis. To assess whether the in vitro expansion of synovium-derived cells affects gene expression, we performed microarray analyses on cells at passage 0, 1, 2, 4, 6, and 8. Synovium-derived cells were rapidly expanded in vitro through passage 8 (about 130 days), and after passage 6, the proliferation rates decreased slightly with a wide range of individual variations. The expressions of CD166, CD49a, and CD106 decreased, whereas those of CD10, CD29, CD44, CD73, CD90, and CD105 showed no significant change. Karyotype analysis revealed no evidence of chromosome abnormalities. The staining of type II collagen and GAG in differentiated cell pellets showed rapid weakening. Genome-wide microarray analysis showed that synovium-derived cells from late passages over-expressed genes associated with cell cycle prolongation and cell aging, and less-expressed genes associated with cell growth stimulation. The in vitro expansion of synovium-derived cells was accompanied with decreased proliferative capacity and the chondrogenic phenotype, which might be modulated by change in gene expression patterns. ß

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

“…), it is important to prepare protocols for an easier and cheaper preliminary phenotype confirmation by means of methods, available in most cell laboratories around the world. Since the desired phenotype can be identified by chondrocyte specific production (Chen et al, 2014; Han et al, 2010), we believe that the easiest and safest preliminary method to prove phenotype preservation could be the analysis of gene expression. More specifically, this analysis should include the evaluation expression of genes related to cartilage specific markers (e.g., collagen type 2 and aggrecan).…”

Applied osmotic loading for promoting development of engineered cartilage

“…SDSCs maintain their chondrogenic capacity regardless of donor age or disease condition (De Bari et al, 2001; Nagase et al, 2008) and have potential for tissue-engineering applications aimed at cartilage repair or regeneration (Bilgen et al, 2007; Han et al, 2010; Lima et al, 2007; Pei et al, 2008a; Pei et al, 2008b). We previously cultured SDSCs in a 3D hydrogel which achieved properties approaching those of native juvenile bovine cartilage.…”

Growth Factor Priming of Synovium-Derived Stem Cells for Cartilage Tissue Engineering

“…Synovium-derived stem cells (SDSCs) have the potential for tissue engineering applications aimed at cartilage repair or regeneration. 5,11,12 A pure population of SDSCs (type B-synovial fibroblasts) can be isolated through multiple passages or by negative isolation, to remove cells that are positive for CD14 (type A-synovial macrophages). 11 The cells can then be expanded in culture for three-dimensional (3D) encapsulation and the expansion media can be optimized to promote chondrogenesis.…”