Generation and characterization of three induced pluripotent stem cell lines for modeling coronary artery vasospasm
DOI: 10.1016/j.scr.2024.103644
Authors: Ines Ross Tacco; Joseph Olshausen; Tse Yuan Chan; Naima Turbes; Ming-Yow Hung; Chi-Tai Yeh; Patricia K. Nguyen; Karim Sallam; Nazish Sayed; Ian Y. Chen
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “adherent” human PSC in “StemMACS iPS-Brew XF”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| Y27632 | 1.0e-5 | mol/l |
| EDTA | 5.0e-7 | mol/l |
Ideally, “PSC” should express “NANOG”, “SOX2”, “OCT3/4” at the end of culture.
Step 2: mesoderm 1
day 1 ~ 3
culture “adherent” human PSC in “RPMI 1640” for “2” days. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| CHIR90021 | 5.0e-6 | mol/l |
| B27 (-Insulin) | — | — |
Step 3: mesoderm 2
day 3 ~ 4
culture “adherent” human PSC in “RPMI 1640” for “1” days. 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 | |
|---|---|---|
| B27 (-Insulin) | — | — |
Ideally, “ME” should express “BRACHYURY”, “TBX6” at the end of culture.