Table 1.
Different Methods Tested for Their Capacity to Sustain Naïve hESCs Derivation
| Method | Blastocyst stage | Number | Medium 1 | PICMI? | Day of outgrowth fragmentation | ROCKi? | Enzyme | Medium 2 |
|---|---|---|---|---|---|---|---|---|
| 1 | Day 6 | 21 | hESC+bFGF | Yes (4/21) | Day 5 | Yes | Trypsin | N2B27+2i+LIF |
| 2 | Day 6 | 15 | 50% N2B27+2i+ LIF+50% hESC+ bFGF |
No | Day 5 | Yes | Trypsin | N2B27+2i+LIF |
| 3 | Day 6 | 13 | N2B27+2i+LIF | No | Day 5 | Yes | Trypsin | Same as medium 1 |
| 4 | Day 5 | 19 | N2B27+2i+LIF | No | Day 5 | No | Trypsin | Same as medium 1 |
| 5 | Day 6 | 14 | N2B27+2i+SB+ LIF |
No | Day 5 | No | Trypsin | Same as medium 1 |
| 6 | Day 6 | 13 | N2B27+2i+Wnt5a+LIF | No | Day 5 | No | Trypsin | Same as medium 1 |
| 7 | Day 5 | 9 | N2B27+SB+PD+ LIF+Epia |
No | Day 3 | No | Accutase | Same as medium 1 |
| 8 | Day 5 | 10 | N2B27+SB+PD+ LIF+Epia+Wnt5a |
No | Day 3 | No | Accutase | Same as medium 1 |
a
Epinephrine was also added during embryo culture, from day 4 to day 5.
hESC, human embryonic stem cells; bFGF, basic fibroblast growth factor; 2i, PD0325901 and CHIR99021; LIF, leukemia inhibitory factor; Epi, epinephrine.