Table 3.
Sample protocols including conditioned medium in the treatment of OA chondrocytes
| MSC source | CM producing | Subject | Experimental conditions | Outcome: CM impact | Ref |
|---|---|---|---|---|---|
| human ASC |
- base: complete serum-free DMEM; - cells seeded 1 × 105 per well; - collection time: after 2 and 7 days (pooled) |
- chondrocytes and synoviocytes from patients with OA K&L = 3 or 4 |
- time: 7 days; - DMEM:CM ratio: 75:25; 50:50; 25:75 |
- no decrease in IL6, MCP-1, MIP1-α, RANTES expression (in contrast with coculture); - no decrease in IL8, GROα expression in synoviocytes; - partially decreased IL8, expression of GROα in chondrocytes; - less efficient reduction in expression of MMP-13, ADAMTS4, ADAMTS5 and increase in the expression of TIMP-1, TIMP-3 (in contrast with coculture); - CM contains a low concentration of PGE2 to reduce inflammation |
[135] |
| human AD-MSC |
- base: DMEM/F12; - cells seeded 2 × 106; - cells at 80–90% confluence |
- chondrocytes and synoviocytes from patients with OA K&L = 4 |
- DMEM/F12 with 1 ml CM; - coculture of chondrocytes and synoviocytes induced with IL-1β |
- reduction of GAGs concentration; - reduction in NO production; - reduction in expression of IL1β, MMP-13, ADAMTS5 in cartilage; - reduction in expression of MMP-13 and ADAMTS5 in synovium; - enhancement in expression of TIMP-1 in cartilage and synovium |
[136] |
| human ASC |
- base: complete serum-free DMEM; - cells after the 4th passage; - cells at between 80 and 90% confluence |
- chondrocytes from patients with OA |
- time: 24 h, 48 h, 72 h; - complete DMEM with CM at a ratio of 1:5; - with/without TNFα stimulation |
- blunting of hypertrophy; - reduction of osteocalcin, COL10, MMP-3, MMP-13 activity; - CM contains chondroprotective factors such as OPG and DKK-1 |
[137] |
| human ASC |
- base: DMEM/F12 with 15% human serum (AB-blood group donors); - cells before and after first passage; - cells at semi-confluence |
- chondrocytes from patients with OA |
- time: 1–7 days; - DMEM/F12 with 10% human serum with/without 1 ml of CM for 6-well plates or 2 ml of CM for 3.5 cm plates; - cells stimulated with IL-1β |
- reduction of SA-β-gal; - reduction of γH2A.X foci; - reduction in formation of actin stress fibres; - reduction in production of oxidative stress and protein kinases; - reduction in expression of caveolin-1 and p21; - reduction of p53 acetylation |
[138] |
| human WJ-MSC |
- base: serum-free DMEM; - cells after the 4th passage; - cells at 70% confluence; - collection time: after 48 h |
- chondrocytes from OA patients |
- time: 3 and 6 days; - DMEM/FBS with WJ-MSC-CM |
- upregulation in expression of COL2A1, ACAN, COMP, and SOX9 after 3 and 6 days in monolayer and mass culture; - reduction in expression of chondrocyte-specific genes after 6 days in monolayer; - maintenance of high expression of chondrocyte-specific genes after 6 days in mass culture |
[49] |
| human WJ-MSC |
- base: complete medium; - cells seeded 3 × 105 per well; - cells treated with IGF1 (0 or 150 ng/ml) for 7 days |
- chondrocytes CHON002 (ATCC CRL-2847) |
- time: 7–14 days; - 15% or 30% CM in medium; - after CM, cells treated with IL-1β for 5 days |
- increase in expression of COL2 in 15% IGF1-WJMSC-CM; - reduction in concentration of inflammatory cytokines; - reduction in expression of MMP-3 |
[47] |
| human SM-MSC |
- base: complete medium; - cells seeded 5 × 105 per flask; - cells at 80% confluence; - cells treated with IGF1 (0 or 150 ng/ml) for 7 days |
- chondrocytes CHON002 (ATCC CRL-2847) |
- time: 7 days; - 15% or 30% CM in medium; - after CM, cells treated with IL-1β for 5 days |
- increase in expression of SOX9, and COL2; - reduction in expression of COL10, MMP-13, ADAMTS4; - IGF1-preconditioning improves the effect of CM in lowering hypertrophic factors and raising anti-hypertrophic factors |
[139] |
| human SHED |
- base: serum-free DMEM/F12; - cells seeded 5 × 103 per cm2 until 70% confluence; - collection time: after 48 h or 72 h |
- chondrocytes from patients with OA |
- time: 48 h; - cells treated with CM collected after 48 h or 72 h; - cells stimulated with IL-1β |
- increase in the concentration of anti-inflammatory factors; - increase in expression of COL2 and ACAN; - decrease in expression of MMP-13 and NF-κB |
[31] |
| human BM-MSC |
- base: serum-free DMEM w/o antibiotics; - cells after the 3rd passage; - cells at 70% confluence; - collection time: after 48 h |
- chondrocytes from rat model of OA (in vivo) |
- time: 8 weeks; - weekly CM injection (100 µl) |
- well-preserved subchondral bone structure; - more abundant cartilage matrix; - reduced MMP-13/TIMP-1 ratio; - inhibited chondrocyte apoptosis with increased autophagy |
[140] |
ACAN aggrecan, ADAMTS- A Disintegrin and Metalloproteinase with Thrombospondin motifs, AD-MSCs adipose tissue-derived MSCs, ASCs adipose stromal cells, BM-MSCs bone marrow MSCs, CM conditioned media, COL10 collagen type X, COL2A1 collagen type II α 1 chain, COMP cartilage oligomeric matrix protein, DKK-1 dickkopf 1, DMEM Dulbecco's Modified Eagle Medium, DMEM/F12 Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12, GAGs glycosaminoglycans, GROα growth-regulated α protein, IGF-1 insulin-like growth factor 1, IL-1β /-6 /-8 interleukin 1β /6 /8, K&L Kellgren and Lawrence, MCP-1 monocyte chemotactic protein 1, MIP1-α macrophage inflammatory protein 1α, MMPs matrix metalloproteinases, MSC mesenchymal stromal cell, NF-κB nuclear factor kappa-light-chain-enhancer of activated B cells, NO nitric oxide, OA osteoarthritis, OPG osteoprotegerin, p21 /p53 protein 21 /53, PGE2 prostaglandin-E2, RANTES regulated upon activation, normal T cell expressed and presumably secreted, SA-β-gal senescence-associated β-galactosidase, SHED human exfoliated deciduous teeth stromal cells, SM-MSCs synovial membrane MSCs, SOX9 SRY-Box transcription factor 9, TIMP-1 /-2 tissue inhibitor of metalloproteinases-1 /-2, TNFα tumour necrosis factor α, WJ-MSCs Wharton's jelly MSCs