Table 2.
Aggregation Enhances MSC Differentiation Potential Compared to 2D Monolayer
| MSC source | Method to generate aggregates | Cell organization | Differentiation potential | Ref. |
|---|---|---|---|---|
| Osteogenic differentiation | ||||
| Human bone marrow | Pellet culture | Aggregates | Qualitative Alzarin red, collagen type I, and bone sialoprotein staining (no comparison with 2D) | 77 |
| Human bone marrow | Modified AggreWell™ | Aggregates | 2-fold increase in ALP, 2- to 5-fold increase in osteocalcin, osteopontin, ALP, and RUNX2, 14-fold increase collagen I | 89 |
| Human bone marrow | Low attachment plates | Aggregates | Two to eightfold increase in osteocalcin, osteopontin | 6 |
| Mouse bone marrow | AggreWell | Dissociated aggregates | Fourfold increase in Alzarin red staining | 35 |
| Rabbit bone marrow | 3D cell sheet | 3D cell sheet | Twofold increase in ALP, osteocalcin, osteopontin, collagen type I | 141 |
| Human adipose tissue | PEG micropatterned surface | Aggregates | Qualitative Alzarin red, fourfold increase in RUNX2 | 34 |
| Human adipose tissue | Chitosan membrane | Aggregates | Qualitative Alzarin red, two to threefold increase in RUNX2 | 71 |
| Rhesus monkey bone marrow | Hanging drop | Dissociated aggregates | 25% increase in ALP, twofold increase calcification and 20% increase in osteocalcin expression | 78 |
| Chondrogenic differentiation | ||||
| Rabbit and human bone marrow | Pellet culture | Aggregates | Collagen type II expression (no comparison with 2D) | 75,77 |
| Human synovial membrane | Hanging drop and pellet culture | Aggregates | 10%–20% increase in collagen type II, aggrecan, and SOX9 of hanging drop compared to pellet cultures | 22 |
| Adipogenic differentiation | ||||
| Mouse bone marrow | Forced aggregation | Dissociated aggregates | Threefold increase in Oil red O staining | 35 |
| Human bone marrow | Low attachment plates | Aggregates | Qualitative Oil red O staining, two to threefold increase in PPAR-γ, LPL, C/EBP-α, and aP2 | 6 |
| Human adipose tissue | PEG micropatterned surface | Aggregates | Qualitative Nile Red staining, increase PPAR-γ expression (14-fold), increased expression of LRP2, FABP4, LPL | 34 |
| Human adipose tissue | Nanoculture plates | Aggregates | Twofold increase in triglycerides accumulation | 73 |
| Human gingiva | Low attachment plates | Aggregates | Increased Oil Red O staining | 12 |
| Rhesus monkey adipose tissue | Hanging drop | Dissociated aggregates | 10%–15% increase in Oil red O and LPL staining | 78 |
ALP, alkaline phosphatase; RUNX2, Runt-related transcription factor 2; PPAR-γ, peroxisome proliferator-activated receptor gamma; LPL, lipoprotein lipase; C/EBP-α, CAAT/enhancer binding protein α; aP2, adipocyte protein 2; LRP2, low-density lipoprotein-related protein 2; FABP4, fatty acid binding protein 4; 3D, three-dimensional.