Successful isolation and ex vivo expansion of human mesenchymal stem/stromal cells obtained from different synovial...

Mesenchymal stromal cells (MSC) isolated from synovial tissues constitute a novel source of stem-like cells with promising applications in cartilage regeneration and potentially in other regenerative medicine and tissue-engineering settings. Detailed characterization of these cells is lacking, thus compromising their full potential. Here we present the detailed characterization of the ex vivo expansion of synovium-derived stromal cells collected by three different biopsy methods: synovium-direct biopsy, arthroscopic trocar shaver blade filtrate, and cells isolated from synovial fluid (SF) samples. Isolation success rates were >75% for all sources. MSC obtained from the different samples displayed the characteristic immunophenotype of adult MSC, expressing CD73, CD90, and CD105. Arthroscopic shaver blade-derived cells showed the higher proliferation capacity measured by the fold increase (FI) in total cell number over several passages and considering their cumulative population doublings (CPD; 15 ± 0.85 vs. 13 ± 0.73 for synovium vs. 11 ± 0.97 for SF). Also, these cells were able to sustain an increased proliferation under hypoxic (2% O₂ ) conditions (FI 55 ± 4 vs. 37 ± 7) after 17 days in culture. Expanded cells were able to differentiate successfully along the osteogenic, adipogenic, and chondrogenic lineages in vitro. Overall, these results demonstrate that synovial tissues represent a promising source for the isolation of human MSC, while depicting the variability associated to the biopsy method used, which impact cell behavior in vitro.