Mesp1 Marked Cardiac Progenitor Cells Repair Infarcted Mouse Hearts
DOI: 10.1038/srep31457
Authors: Yu Liu; Li Chen; Andrea Diaz Diaz; Ashley Benham; Xueping Xu; Cori S. Wijaya; Faisal Fa’ak; Weijia Luo; Benjamin Soibam; Alon Azares; Wei Yu; Qiongying Lyu; M. David Stewart; Preethi Gunaratne; Austin Cooney; Bradley K. McConnell; Robert J. Schwartz
Step 1: iPSC/ESC maintainance
day 0 ~ 1 (duration missing in database; assumed 1 day)
culture “unspecified method” mouse ESC in “DMEM”. Medium should contain factors shown in the following table.
| concentration | unit | |
|---|---|---|
| FBS | 15 | % |
| sodium pyruvate | 0.001 | mol/l |
| NEAA | 1 | % |
| LIF | 1.0e+6 | U/l |
| 2-mercaptoethanol | 1.0e-4 | mol/l |
| PSA | 50000 | U/l |
| streptomycin | 0.05 | g/l |
Ideally, “ESC” should be obtained at the end of culture.
Step 2: mesoderm 1
day 1 ~ 3
culture “spheroid” mouse ESC in “IMDM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| F12 | 25 | % |
| BSA | 0.05 | % |
| penicillin G | 100000 | U/l |
| streptomycin | 0.1 | g/l |
| L-glutamate | 0.002 | mol/l |
| chemically defined lipids | 1 | x |
| MTG | 4.5e-4 | mol/l |
| B27 | 0.5 | x |
| N2 | 1 | x |
| PVA | 0.4 | g/l |
| Insulin | 0.01 | g/l |
| Y27632 | 1.0e-6 | mol/l |
| BMP4 | 2.0e-5 | g/l |
Step 3: mesoderm 2
day 3 ~ 5
culture “spheroid” mouse ESC in “IMDM” for “2” days. Medium should contain factors shown in the following table, and should be changed once in “2” days.
| concentration | unit | |
|---|---|---|
| F12 | 25 | % |
| BSA | 0.05 | % |
| penicillin G | 100000 | U/l |
| streptomycin | 0.1 | g/l |
| L-glutamate | 0.002 | mol/l |
| chemically defined lipids | 1 | x |
| MTG | 4.5e-4 | mol/l |
| B27 | 0.5 | x |
| N2 | 1 | x |
| Y27632 | 1.0e-6 | mol/l |
| BMP4 | 2.0e-5 | g/l |
| IWR1 | 5.0e-6 | mol/l |
| SB431542 | 1.0e-5 | mol/l |
Ideally, “EC” should be obtained at the end of culture.