Materials.
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 15 cm dishes | Corning | 430599 | tissue culture treated |
| 35 mm dishes | Corning | 430165 | tissue culture treated |
| 6-well plates | Corning | 3516 | tissue culture treated |
| Antimicrobial reagent Brand Name: Normocin | Invitrogen | ant-nr-1 | |
| bFGF | R&D systems | 233-FB-025 | use at 50 ng/mL |
| BMP4 | BioLegend | 595202 | use at 25 ng/mL |
| Bovine Serum Albumin (BSA) | Fisher Scientific | BP1600–1 | |
| Cell Detatchment Solution Brand Name: Accutase | Stemcell Technologies | 7920 | |
| Dimethyl Sulfoxide (DMSO) | Sigma Aldrich | D2650–100mL | |
| Dispase | Stemcell Technologies | 7913 | |
| DLL4 | R&D systems | 1506-D4/CF | recombinant human; use at 10 μg/mL |
| DMEM:F12 | Gibco | 11320–033 | |
| Dulbecco’s Phosphate Buffered Saline (PBS) | Gibco | 14190144 | |
| Endothelial cell growth medium Brand Name: EGM-2 Endothelial Cell Growth Medium-2 BulletKit (EGM-2) | Lonza | CC-3162 | |
| FACS tubes | Corning | 352235 | polystyrene round bottom with filter cap |
| Fetal Bovine Serum (FBS) | Gemini Bio | 100–106 | |
| Fibronectin | ThermoFisher Scientific | 33016015 | use at 4 μg/cm2 |
| GSK3i/CHIR99021 | Stemgent | 04–0004–02 | 10 mM stock; use at 5 μM |
| Hanks Balanced Salt Solution (HBSS) | Gibco | 14175–095 | |
| Hydrochloric Acid (HCl) | Fisher Scientific | A144S-500 | |
| Matrix protein Brand Name: Matrigel | Corning | 356230 | Growth factor reduced. Refer to the Certificate of Analysis for the lot to determine the protein (Matrigel) concentration. This concentration is required to calculate the volume of Matrigel that contains 1 mg of protein. |
| Methylcellulose-based medium Brand Name: MethoCult H4435 Enriched | Stemcell Technologies | 4435 | |
| Pluripotent stem cell differentiation medium Brand Name: STEMdiff APEL 2 | Stemcell Technologies | 5270 | |
| Pluripotent stem cells: H1, H9, H9-FUCCI | WiCell | WA09 (H9), WA01 (H1) | human; H9-FUCCI were obtained from Dr. Ludovic Vallier’s lab at Cambridge Stem Cell Institute |
| Protein-Free Hybridoma Medium (PFMH) | Gibco | 12040077 | |
| Retinoic Acid | Sigma Aldrich | R2625–50mg | use at 0.5 μM |
| Reverse transcription master mix Brand Name: iScript Reverse Transcription Supermix | BioRad | 1708840 | |
| RNA extraction kit Brand Name: RNeasy Mini Kit | Qiagen | 74104 | |
| Sodium Hydroxide (NaOH) | Fisher Scientific | SS255–1 | |
| Stem cell growth medium Brand Name: mTeSR1 | Stemcell Technologies | 85850 | |
| SYBR Green master mix Brand Name: iTaq Universal SYBR Green Master Mix | BioRad | 1725121 | |
| Trypsin-EDTA | Gibco | 25299956 | 0.25% |
| VEGF165 (VEGF-A) | PeproTech | 100–20 | use at 50 ng/mL |
| α-CD31-FITC | BioLegend | 303104 | 2 μg/mL* |
| α-CD45-APC/Cy7 | BioLegend | 304014 | 2 μg/mL* |
| α-Flk-1-PE/Cy7 | BioLegend | 359911 | 2 μg/mL* |
| α-c-Kit-APC | BioLegend | 313206 | 2 μg/mL* |
| α-VE-Cadherin-PE | BioLegend | 348506 | 2 μg/mL* |
| α-CD34-Pacific Blue | BioLegend | 343512 | 2 μg/mL* |
*
Antibody fluorescent conjugates should be optimized based on the cell sorter used. Presented here are the final concentrations utilized in this study.