Table 1.
Fabrication Methods and the Properties of MSC Aggregates
| Methods | MSC source | Starting cell density | Initial cell number per aggregate | Initial aggregate size | Aggregate size after long-term culture | Long-term viability | Proliferation | Ref. |
|---|---|---|---|---|---|---|---|---|
| Nonadhesive 96-well plate | Human bone marrow | 1×105 cells/mL | 1×105 | 632±35 μm | 353±28 μm at day 21 | +++ (at day 17) | No proliferation during a 21-day culture period | 50 |
| Nonadhesive 96-well plate | Human umbilical cord | 1×104–1.8×105 cells/mL | 1×104–1.8×105 | 150 μm to 1.0 mm | ND | ND | ND | 36 |
| Low attachment plate | Human umbilical cord | ND | ND | 40–100 μm | ND | ND | Increased proliferation compared to 2D at day 1 (cell cycle analysis) | 59 |
| Low attachment plate | Human gingiva | 2×105 cells/mL | 20–100 | 20–100 μm | ND | ++ (at day 3) | ND | 12 |
| Low attachment plate+agitation | Human bone marrow | 2×104 cells/mL | ND | 32–100 μm (day 7) | No change in bioreactor cultures | ++ (at day 7) | Similar to monolayer at day 7 (cell cycle analysis) | 6 |
| Hanging drop+static semisolid | Human bone marrow | 5×103 cells/mL | ND | ND | 159±19 μm at day 17 | ++ (at day 17) | ND | 50 |
| Polypropylene tube+suspension | Human bone marrow | 2×104 cells/mL | 2×104 | ND | 360±60 μm at day 17 | ++ (at day 17) | ND | 50 |
| Thermal lifting | Human bone marrow | 3×103 cells/cm2 | 10 | 89±32 μm | 100–250 μm at day 2 | ++ | Increased proliferation compared to self-assembly (BrdU assay at day 1) | 49 |
| Forced aggregation | Human bone marrow | 2.5×105 cells | 2.5×105 | ±1 mm after 1 week | No change after 4 weeks | ++ (at day 21) | No proliferation after 7 days (cell cycle analysis) | 77 |
| Forced aggregation +agitation | Mouse bone marrow | 1.8–6×106 cells/mL | 300–1000 | 100 μm (day 7) | No change in stirred culture at day 7 | ++ (up to day 7) | Similar to monolayer (BrdU assay at day 7) | 35 |
| PEG-micropatterned surface | Human bone marrow and UET-13 line | 5×104 cells/cm2 | 50 | 100 μm | ND | + (day 5–15) | Decreased proliferation compared to 2D | 34,73 |
| Chitoson membrane | Human adipose tissue | 1×103–5×104 cells/cm2 | ND | 22–54 μm | Increased size, 77–195 μm day 7 | + | No proliferation after 7 days (Alamar blue) | 71,72 |
MSC, mesenchymal stem cell; PEG, poly(ethylene glycol); BrDU, 5-bromo-2-deoxyuridine; 2D, two-dimensional; ND, not determined.