TABLE 1.
Comparison of different studies reporting static and dynamic methods of spheroid formation and their results for the dynamic cultivation of MSC spheroids.
| Ref. | MSCs source | Ps | Formation method | Dynamic culture | T° | rpm | O2 | Medium | Time | Initial cell density | Priming | Cells per spheroid | Spheroid parameters |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Frith et al. (2010) | human BM-MSCs | nr. | Static (ex situ) Microwells in non-adherent plates | Small- and large-scale SF and RWV | 37°C | 30 (SF) and 15 (RWV) | nr. 21% ia. | αMEM + 15% FBS + pen/strep | 7 days | 1 × 106 cells/ml (formation) 2 × 104 cells/ml (culture) | nr. | Random | Diameter 98.7 (SF) and 31.7 µm (RWV) |
| Bhang et al. (2011) | human AD-MSCs | 5 | Dynamic (in situ) Single-cell inoculation in SF | Small-scale: SF | 37°C | 70 | 1 and 5% (culture) | αMEM + 10% FBS + pen/strep | 3 days | 6 × 105 cells/ml | nr. | Random | Diameter 100–350 µm |
| Hildebrandt et al. (2011) | human BM-MSCs | 3 to 10 | Dynamic (in situ) Non-adherent bacterial plate on a rotating platform | Small-scale: Bacterial culture dishes on a rotating platform | 37°C | 75 for 24 h then raised to 85 to 95 | nr. 21% ia. | αMEM + 15% FBS + pen/strep | 21 days | 5 × 104 cells/ml | nr. | Random | Multiaggregation (diameter not reported) |
| Hildebrandt et al. (2011) | human BM-MSCs | 3 to 10 | Static (ex situ) HP | Small-scale: Bacterial culture dishes on a rotating platform | 37°C | 75 for 24 h then raised to 85 to 95 | nr. 21% ia. | αMEM + 15% FBS + pen/strep | 21 days | 5 × 103 cells per drop (formation) | nr. | 5,000 | Multiaggregation (diameter not reported) |
| Hildebrandt et al. (2011) | human BM-MSCs | 3 to 10 | Dynamic (in situ) Non-adherent bacterial plate on a rotating platform | Small-scale: Plate on a rotating platform | 37°C | 75 for 24 h then raised to 85 to 95 | nr. 21% ia. | αMEM + 15% FBS + pen/strep | 21 days | 6 × 104 cells per well | nr. | 10000/15000/20000 | Diameter 267, 382 and 435 µm |
| Baraniak & McDevitt, (2012) | Murine BM-MSCs | 8 to 10 | Static (ex situ) Microwells non-adherent plates | Small-scale: Bacterial culture dishes on a rotating platform | 37°C | 45 | nr. 21% ia. | CEM + 10% FBS + 10% HS + 2 mM l-glutamine + pen/strep/ampho | 21 days | 1.8–6 × 106 cells/ml (static) and 1.500 spheroids in 10 ml (dynamic) | nr. | 300/600/1,000 | Area 19.3, 24.7, and 7.9 × 103 μm2 by day 7 |
| Bhang et al. (2012) | human CB-MSCs | * | Static (ex situ) HP | Small-scale: SF | 37°C | 70 | 21 and 1% | αMEM + 10% FBS | 2 days | 1 × 104-2 × 106 cells/ml (static) and 6 × 105 cells/ml (dynamic) | nr. | 300/60000 | Diameter 200–400 µm |
| Cho et al. (2012) | human AD-MSCs | >5 | Static (ex situ) HP | Small-scale: SF | nr. 37°C ia. | 70 | 21 and 1% O2. | αMEM + 10% FBS + pen/strep | 2 days | 1 × 106 cells/ml l (static) and 4.2 × 107 cells/ml (dynamic) | nr. | 30,000 app. | nr. |
| Alimperti et al. (2014) | human BM-MSCs | 4 | Dynamic (in situ) Single cell inoculation in SF | Small-scale: SF | nr. 37°C ia. | 80 | 21% O2 | Serum-free medium | 7 days | 1 × 105 cells/ml | nr. | Random | nr. |
| Bhang et al. (2014) | human AD-MSCs | up to 5 | Static (ex situ) HP | Small-scale: SF | 37°C | 45 | 21 and 1% | αMEM or CRM | 4 days | 30 µl drople with 1 × 106 cells/ml (static) 6 × 105 cells/ml (dynamic) | nr. | 30.000 app. | nr. |
| Zimmermann and Mcdevitt, (2014) | human BM-MSCs | 4 | Static (ex situ) Microwells in non-adherent plates | Small-scale: Bacterial culture dishes on a rotating platform | 37°C | 65 | nr. 21% O2 ia. | Serum-free MesenCult-XF.IFN-y and TNFα were added on day 1 | 4 days | 1.2, 3.0, and 6.0 × 106 cells/ml (formation) and 3 × 106 cells/ml (dynamic) | Yes. IFN-ϒ and TNF-α. | 200/500/1,000 | nr. |
| Kwon et al. (2015) | human AD-MSCs | 5 | Dynamic (in situ) Single-cell inoculation in SF | Small-scale: SF | 37°C | 70 | nr. 21% ia. | αMEM + 10% FBS + pen/strep | 5 days | 1 × 106 cells/ml | nr. | Random | nr. |
| Li et al. (2015) | human UC-MSCs | 1 | Dynamic (in situ) Single-cell inoculation in dishes on a rocker system | Small-scale: Bacterial culture dishes on a rocker system | 37°C | 10 | nr. 21% ia. | Serum-Free Medium Bao et al. (2013) | 9 days | 1 × 106 cells/mI | nr. | Random | Diameter of 118 μm at day 9 |
| Santos et al. (2015) | human UC-MSCs | 3 to 12 | Dynamic (in situ) Single-cell inoculation in SF | Small-scale: SF | 37°C | 80 (formation) 110 (culture) | nr. 21% ia. | MEM +2 mM l-glutamine + 1 g/L glucose + 2.2 g/L sodium bicarbonate + 10% FBS | 11 days | 1 × 106 cells/ml | nr. | Random | Diameter of 143 μm at day 2 and 309 μm from day 4–11. |
| Zhang et al. (2015) | human AD-MSCs | 5 | Dynamic (in situ) RWV | Large-scale: RWV | 37°C | 25 | nr. 21% ia. | DMEM/F12 + 10%(v/v) FBS + 1% pen/strep | 5 days | 1 × 106 cells/ml | nr. | Random | Diameter of 123 μm at day 5 |
| Costa et al. (2017) | human BM-MSCs | 4 to 6 | Static (ex situ) Microwells in non-adherent plates | Small-scale: Ultralow attachment plate on a rotatory orbital shaker | 37°C | 65 | nr. 21% ia. | DMEM + 10% FBS+ 1% antibiotics-antimycotics | 7 days | 5 × 104 cells/ml | nr. | 500 | Area of 56400 μm2 at day 7 |
| Egger et al. (2017) | human AD-MSCs | 2 | Dynamic (in situ) Single-cell inoculation on STR | Large-scale: STR | 37°C | 600 | 21 and 5% O2 | αMEM + 0.5% gentamycin + 10% human platelet lysate + 1 U/ml heparin | 6 days | 1 × 105 cells/ml | nr. | Random | nr. |
| Cha et al. (2018) | human BM-MSCs | 2 | Static (ex situ) Microwells in non-adherent plates | Small-scale: Plates in an orbital shaker | 37°C | 30 | nr. 21% ia. | DMEM +10% FBS or exosome-free FBS + 1% antibiotics-antimycotics | 7 days | 400 cells per well (formation) | nr. | 400 app. | Diameter of 150 µm (post-formation) |
| He et al. (2019) | rabbit BM-MSCs | 5 | Dynamic (in situ) Single-cell inoculation on SF | Small-scale: SF | nr. 37°C ia | 40/45/50 | nr. 21% ia. | αMEM + 10% FBS + pen/strep | 5 days | 2,4 and 8 × 105 cells/ml (at 50 rpm) | nr. | Random | Diameter of 40–60 µm |
| Allen et al. (2019) | human SyF-MSCs | 5 | Static (ex situ) Microwells in non-adherent plates | Large-scale: STR | 37°C | 80 | nr. 21% ia. | PPRF-msc6 serum-free | 12 days | 5 × 105 cells/ml | nr. | 500/1000/1,500/2000 | Diameter of 121, 145, 161 and 181 μm (spheroid formation) and 124 μm on day 1 and increased to 643 μm by day 12 (dynamic culture) |
| Allen et al. (2019) | human SyF-MSCs | 5 | Dynamic (in situ) Single-cell inoculation in a STR | Large-scale: STR | 37°C | 80 | nr. 21% ia. | PPRF-msc6 serum-free | 12 days | 5 × 105 cells/ml | nr. | Random | Diameter of 100 μm at day 6, 125 μm on day 8, 137μm on day 10, and 153 μm on day 12. |
| Miranda et al. (2019) | human UC-MSCs | 3 to 12 | Dynamic (in situ) Single-cell inoculation in SF | Small-scale: SF | 37°C | 80 (formation) and 110 (culture) | nr. 21% ia. | αMEM + 15% FBS | 7 days | 1 × 106 cells/ml (dynamic) | nr. | Random | Diameter of 195.48 µm from day 5–7 of culture |
| Niibe et al. (2020) | human BM-MSCs | 4 to 7 | Dynamic (in situ) Single-cell inoculation in SF | Small-scale: SF | 37°C | 85–95 | nr. 21% ia. | αMEM, GlutaMAX-I, 10% FBS, 1% p/s, 10 mM hepes, 20 ng/ml FGF2. | 1–2 months | 5 × 105–5 × 106 cells/ml | nr. | Random | Feret’s diameter 5 × 105: 699 µm (1) 757 µm (2) 5 × 106: 782 µm (1) 833 µm (2) |
| Niibe et al. (2020) | human BM-MSCs | 17 to 20 | Dynamic (in situ) Single-cell inoculation in SF | Small-scale: SF | 37°C | 85–95 | nr. 21% ia. | αMEM, GlutaMAX-I, 10% FBS, 1% p/s, 10 mM hepes, 20 ng/ml FGF2. | 1–2 months | 5 × 105–5 × 106 cells/ml | nr. | Random | Feret’s diameter 5 × 105: 534 µm (1) 734 µm (2), 5 × 106: 760 µm (1) 1,158 µm (2) |