Table 1.
Comparison of the culture techniques, generation time, and plating densities of organoids
| Organoid | Origin | No. Plated cells/tissues | Method/Comments | Time to generation (days) | Ref. |
|---|---|---|---|---|---|
| Anterior | |||||
| Cortical Neurones | m/hESCs | 3 × 103 cells | SFEBq; low‐adhesion U‐bottomed plates | 10–20 | 27 |
| Optic cup | mESCs | 3 × 103 cells | SFEBq, Matrigel embedding; low‐adhesion U‐bottomed plates | ∼9 | 26 |
| hESCs | 9 × 103 cells | SFEBq low‐adhesion V‐bottomed plates; Matrigel embedding day 2; transfer to petridish day 12; | ∼24 | 10 | |
| SFEBq; low‐adhesion plates; Matrigel embedding | 14–24 | ||||
| Inner Ear | mESCs | 3 × 103 cells | 93, 94 | ||
| Cerebral | mESCs | 2 × 103 cells | EBs generated in low‐adhesion U‐bottomed plates; embedded in Matrigel and cultured in spinning bioreactor | 30–75 | 39 |
| hESCs/iPSCs | 4.5 × 103 cells | ||||
| Neural Tube | mESCs | 1 cell | Cells (5 × 104 cells) in N2B27 embedded in Matrigel, spread evenly over glass‐bottomed MatTek dishes; organoids form from single cells | <10 | 83 |
| The viscera | |||||
| Intestine | Crypts (m) | 500 Crypts | Matrigel embedding; Single LGR5+ forms organoids; enhanced with Paneth cell co‐culture | 8–14 | 11, 47 |
| LGR5+ SC (m) | 1 cell | ||||
| LGR5+ + Paneth (m) | 500 cells each | ||||
| hESCs/iPSCs | 50 spheroids | Monolayer differentiation towards hindgut; formed spheroids embedded in Matrigel | 14–28 | 51, 52 | |
| Colon | Crypts (m, h) | 500 Crypts | Matrigel Embedding; single LGR5+ SCs can form organoids if anoikis is inhibited in first 2 days | 7–10 | 48 |
| LGR5+ SC (m) | 1 × 103 cells | ||||
| Stomach | Gastric glands (m) | 100 glands | Matrigel embedding | 7–10 | 49 |
| LGR5+ SC (m) | 50 cells | ||||
| hESCs/iPSCs | 50 spheroids | Monolayer differentiation towards posterior foregut; spheroids embedded in Matrigel | 28 | 57 | |
| Lung | hESCs | 50 spheroids | Monolayer differentiation towards anterior foregut; spheroids embedded in Matrigel | 65–110 | 53 |
| Kidney | hESCs | 1 × 106 cells | Monolayer differentiation towards intermediate mesoderm, dissociation and culture in Air‐Liquid interface after 18 days in culture | 4 | 43 |
| Liver | hiPSCs | 1 × 106 cells | Monolayer differentiation towards endoderm; co‐culture with HUVECs and hMSCs on Matrigel | 4–6 | 46 |
| Biliary Ducts (m) | 100 glands | Matrigel Embedding | 7 | 95 | |
| LGR5+ SCs (m) | 1 cell | Matrigel Embedding | 19 | 95 | |
| Embryo | |||||
| ‘Gastruloids’ | mESCs | ∼400 cells | Cells plated in low‐adhesion, U‐bottomed plates | 4–5 | 73, 74, 96 |
| Other | |||||
| Skin | Primary keratinocytes (h) | 3 × 105 cells | Air‐liquid interface culture | ∼ 21 | 19 |
| Oesophagus | Oesophageal fibroblasts (m/h) | 2.5 × 105 cells | Fibroblasts embedded in collagen/matrigel; Oesophageal keratinocytes (4 × 105) added after seven days; Air‐liquid interface culture | 11–13 | 18 |
| Blood vessels | HUVECs | 4.5 × 104 cells | Cells seeded into collagen microvessels (mechanical support) | 7–14 | 20 |
Information on selected organoids from a number tissues, broadly grouped into three categories, is given for the tissue origin (e.g. ESC, adult SC, tissue fragments etc.), number of cells or individual cellular units (crypts or cell spheroids) used to generate the organoid and the time taken to form the organoid structure. Time to formation is taken as the amount of time forming organoid structure, not the total time in culture. This is particularly important in the case of the visceral organoids from SCs, where cells are first directed to specific lineages before Matrigel embedding and organoid formation. ESCs: embryonic stem cells; SCs: stem cells; (h): human; (m): mouse; SFEBq: serum‐free floating culture of embryoid‐body‐like aggregates with quick reaggregation; HUVECs: Human Umbilical Vein Endothelial Cells; hMSCs: human Mesenchymal Stem Cells.