Generation of a doxycycline-inducible ETV2 expression cell line using PiggyBac transposase system
DOI: 10.1016/j.scr.2022.102985
Authors: Meng Zhou; Jie Ni; Pufeng Huang; Xujie Liu
Step 1: iPSC/ESC maintainance
day 0 ~ 1
culture “adherent” human ESC in “PGM1 medium” for “1” days. “ESC” should be “unspecified regulation” for “ETV2”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.
| concentration | unit | |
|---|---|---|
| Y27632 | 1.0e-5 | mol/l |
| doxycycline | 0.002 | g/l |
| Puromycin | 3.0e-4 | g/l |
| Puromycin | 0.001 | g/l |
| EDTA | 5.0e-4 | mol/l |
Ideally, “ESC” should express “NANOG”, “POU5F1”, “DPPA4”, “REX1”, “SOX2”, “ETV2” at the end of culture.