Defined MicroRNAs Induce Aspects of Maturation in Mouse and Human Embryonic-Stem-Cell-Derived Cardiomyocytes
DOI: 10.1016/j.celrep.2015.08.042
Authors: Desy S. Lee; Jyh-Hong Chen; David J. Lundy; Chung-Hung Liu; Shiaw-Min Hwang; Lil Pabon; Ru-Chi Shieh; Chien-Chang Chen; Sheng-Nan Wu; Yu-Ting Yan; Sho-Tone Lee; Po-Min Chiang; Shu Chien; Charles E. Murry; Patrick C.H. Hsieh
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “adherent” human ESC in “MEF-conditioned medium”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| FGF | 5.0e-6 | g/l |
Ideally, “ESC” should be obtained at the end of culture.
Step 2: Primitive streak 1
day 1 ~ 2
culture “adherent” human ESC in “RPMI” 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 | — | — |
| Activin A | 1.0e-4 | g/l |
Step 3: mesoderm 1
day 2 ~ 4
culture “adherent” human ESC in “RPMI” 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 | |
|---|---|---|
| B27 | — | — |
| BMP4 | — | — |
| CHIR90021 | — | — |
Step 4: mesoderm 2
day 4 ~ 5 (duration missing in database; assumed 1 day)
culture “adherent” human ESC in “RPMI”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| B27 | — | — |
| XAV939 | — | — |