Skeletal-muscle-derived mesenchymal stem/stromal cells from patients with osteoarthritis show superior biological properties compared to bone-derived cells

Čamernik, Klemen; Mihelič, Anže; Mihalič, René; Presen, Darja Marolt; Janež, Andrej; Trebše, Rihard; Marc, Janja; Zupan, Janja

doi:10.1016/j.scr.2019.101465

Cited by 29 publications

(37 citation statements)

References 36 publications

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“…The skeletal muscle and trabecular bone were harvested from each patient and from all of the donor groups at the same anatomical site: gluteus medius and femoral head, as described previously [18]. Both biopsy samples were immediately placed into growth medium: low-glucose Dulbecco's modified Eagle's medium (DMEM; Biowest), with 10% fetal bovine serum (Gibco), 2-mM L-glutamine (Biowest), and 2% antibiotics/antimycotics (100× stock: 8.5 g/L NaCl, 0.025 g/L amphotericin B, 6.028 g/L penicillin G, sodium salt, 10 g/L streptomycin sulfate; all Biowest).…”

Section: Donor Inclusion and Bone Samplingmentioning

confidence: 99%

“…Flow cytometry was performed on an aliquot of both freshly isolated cells and on the culture-expanded MSCs between passages 1 and 5 (p1-p5), as described previously [18,21]. Freshly isolated cells were tested for CD45, CD19, CD14, and CD34 (mesenchymal fraction) using monoclonal antibodies 2D, SJ25C1, 61D3 (all ThermoFisher Scientific), and AC136 (Miltenyi Biotec), respectively.…”

Section: Immunophenotypingmentioning

confidence: 99%

“…The colony-forming unit fibroblast (CFU-F) assay was performed at p0 and p1, as described previously [18]. Briefly, the CFU-F assay data at p0 were calculated as methyl violet-positive colonies per viable MSCs counted after p0 (%).…”

Section: Colony-forming Unit Fibroblast Assaymentioning

confidence: 99%

“…Senescence analysis was assessed as described previously [18]. Briefly, the cells were seeded as three to five replicates in 12-well plates at 5000 cells/cm 2 .…”

Section: Senescence Analysismentioning

confidence: 99%

“…Multilineage differentiation was performed as described previously [18]. Briefly, for adipogenesis and osteogenesis, the cells were seeded as four replicates in 24-well plates at a density of 25,000 cells/cm 2 .…”

Section: Multilineage Differentiationmentioning

confidence: 99%

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Comprehensive analysis of skeletal muscle- and bone-derived mesenchymal stem/stromal cells in patients with osteoarthritis and femoral neck fracture

et al. 2020

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Background: Mesenchymal stem/stromal cells (MSCs) can replenish the aged cells of the musculoskeletal system in adult life. Stem cell exhaustion and decrease in their regenerative potential have been suggested to be hallmarks of aging. Here, we investigated whether muscle-and bone-derived MSCs of patients with osteoarthritis and osteoporosis are affected by this exhaustion, compared to healthy donors. Methods: Patients with primary osteoarthritis, femoral neck fractures due to osteoporosis, and healthy donors (controls) were included. MSCs were isolated from the skeletal muscle and subchondral bone from each patient and compared using ex vivo and in vitro analyses, including immunophenotyping, colony-forming unit fibroblast assays, growth kinetics, cell senescence, multilineage potential, and MSC marker gene expression profiling. Results: Freshly isolated cells from muscle from patients with osteoarthritis showed a lower proportion of CD45/ CD19/CD14/CD34-negative cells compared to patients with osteoporosis and healthy donors. Freshly isolated muscle cells from patients with osteoarthritis and osteoporosis also showed higher clonogenicity compared to healthy donors. MSCs from both tissues of osteoarthritis patients showed significantly reduced osteogenesis and MSCs from the bone also reduced adipogenesis. Chondrogenic pellet diameter was reduced in bone-derived MSCs from both patient groups compared to healthy donors. A significant positive correlation was observed between adipogenesis and CD271 expression in muscle-derived MSCs. CD73 was significantly lower in bone-derived MSCs from osteoarthritis patients, compared to osteoporosis patients. Gene expression profiling showed significantly lower expression of MSC marker gene leptin receptor, LEPR, previously identified as the major source of the bone and adipocytes in the adult bone marrow, in bone-derived MSCs from patients with osteoarthritis in comparison with osteoporotic patients and healthy donors.

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Section: Donor Inclusion and Bone Samplingmentioning

confidence: 99%

Section: Immunophenotypingmentioning

confidence: 99%

Section: Colony-forming Unit Fibroblast Assaymentioning

confidence: 99%

“…Senescence analysis was assessed as described previously [18]. Briefly, the cells were seeded as three to five replicates in 12-well plates at 5000 cells/cm 2 .…”

Section: Senescence Analysismentioning

confidence: 99%

Section: Multilineage Differentiationmentioning

confidence: 99%

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