Table 2.
Cell type | Culture condition | Added factors | Basal medium | Embryo-derived | Culture- derived | Gene expression | X status | Functional tests | Ref. |
---|---|---|---|---|---|---|---|---|---|
Embryonic stem cells (ESCs) |
Feeders/ serum | - | MEFs/ DMEM/ 20% serum | ✓ | Little transcriptional data | ✓ | IVD, teratomas, pre-imp. Chimaeras, GLT | [4, 12] | |
Serum/LIF‡ | 1000 U/ml LIF | GMEM or DMEM/ 10% serum | ✓ | Naïve markers. Heterogeneous lineage markers. | ✓ | IVD, teratomas, pre-imp. Chimaeras, GLT. | [27, 28] | ||
BMP/LIF | 10 ng/ml BMP4 OR 200 ng/ml GDF6 + 1000 U/ml LIF |
N2B27 | ✓ | ✓ | Little transcriptional data. Homogeneous Oct4 | Pre-imp. Chimaeras, GLT. | [29] | ||
2i* | 1 μm PD032/ 3 μM CHIR/ (+/− LIF) | N2B27 | ✓ | ✓ | Homogeneous naïve markers. Reduced lineage markers (rel. to SL). | ✓ | IVD, pre-imp. Chimaeras, GLT. | [91, 97] | |
KOSR/LIF | 1000 U/ml LIF | Knockout DMEM/ 20% KOSR | ✓ | ✓ | Naïve and PrE markers | ? | IVD, pre-imp. Chimaeras, GLT. | [46, 101] | |
Epiblast stem cells (EpiSCs) | F/A‡ | 5-12 ng/ml FGF2 + 10-20 ng/ml Activin | MEFs, Knockout DMEM/ 20% KOSR or “CDM” [207] | ✓ | ✓ | Primed markers. Heterogeneous lineage markers. | ✗ | IVD, teratoma formation, rare pre-imp. Chimaeras, post-imp. Chimaeras. | [3, 17] |
F4-EpiSC | 25 ng/ml FGF4 | DMEM/ 20% serum | ✓ | Homogeneous Oct4. Primed and lineage markers. | ✗ | Teratomas, pre-imp. Chimaeras. | [160] | ||
F/A/XAV* (WiEpiSC) |
5 ng/ml FGF2/ 10 ng/ml Activin/ 10 μM XAV | DMEM/20% KOSR | ✓ | ✓ | Homogeneous pluri and primed markers. Reduced lineage markers (relative to FA). | ? | Post-imp. Chimaeras –bias towards extraembryonic mesoderm. | [156] | |
F/A/IWP2* (WiEpiSC) |
12 ng/ml FGF2, 20 ng/ml Activin, 2 μM IWP2 | Derivation in DMEM/ 18% KOSR/ 2% serum ➔ N2B27 at P2 | ✓ | Pluri markers. Reduced lineage markers (rel. to FA). | ? | Enhanced ESC reversion, pre-imp. Chimaeras. | [127] | ||
XAV/CHIR | 3 μM CHIR/ 2 μM XAV | GMEM/ 10% serum | ✓ | ✓ | Reduced naïve markers (rel. to ESCs). Primed markers. | ✗ | Teratomas formation. | [20] | |
Potential intermediate states | EpiLCs | 12 ng/ml FGF2, 20 ng/ml Activin | N2B27/ 1% KOSR | ✓ | Reduced pluri/naïve markers. Increased primed markers (rel. to ESCs). Reduced lineage markers (rel. to EpiSCs) | ✗ | Efficient PGCLC generation. | [145] | |
Transient Epi state | 10-12 ng/ml FGF2 | N2B27 (+/− 1% KOSR) | ✓ | Decreased Rex1, Increased Fgf5. | ? | In vitro NMPs. | [154, 155] | ||
INTPSC | 12 ng/ml FGF2, 10 ng/ml Activin, 3 μM CHIR | N2B27/ 1% KOSR | ✓ | Intermediate expression of pluri, naïve, primed and lineage markers. | ✓ | Teratoma formation, pre-imp. Chimaeras. | [155] | ||
FAB-SCs | 1 ng/ml FGF2, 50 ng/ml Activin, 0.5 μM BIO, 100 ng/ml LIF blocking antibody | MEFS/ DMEM/ 15% KOSR | ✓ | Pluri markers. Naïve miRNAs. Reduced naïve/germ cell markers (rel. to ESCs). Little/no primed expression. | ? | Not functionally pluripotent | [161] | ||
IESCS | 12 ng/ml Activin | GMEM/ 10% serum | ✓ | Intermediate expression of pluri, naïve, primed and lineage markers. | ? | IVD, teratoma formation – incorporate into pre-imp. Embryos but negative impact on development. | [162] | ||
EPL | +/− 1000 U/ml LIF | 50% MEDII conditioned medium/ 50% DMEM/ 10% serum | ✓ | Pluri markers. Reduced naïve and increased primed markers (rel. to ESCs). | ? | Not functionally pluripotent |
X status refers to the activation status of the X chromosome, ✗ = inactive X chromosome, ✓ = active X chromosome. DR downregulated, UR upregulated, Epi epiblast, PrE primitive endoderm, IVD in vitro differentiation, pre-imp. pre-implantation, post-imp. post-implantation, GLT germline transmission, pluri pluripotency genes. ‡ = standard culture condition, * = “ground state” of self-renewal, rel. to = relative to