μ-Slide Angiogenesis |
Ibidi |
N/A |
A flat, glass bottom tissue-culture plate with side walls enables facile confocal imaging |
96 well, round bottom, ultra low attachment microplate, sterile |
Corning |
7007 |
Prevents the binding of cell-laden collagen hydrogels to the cell culture dish |
Accutase |
STEMCELL Technologies |
7920 |
Gentle cell detachment solution; does not degrade extracellular epitopes vital for FACS |
Advanced DMEM/F12 |
Thermo Scientific |
12634010 |
The base media for iPSC-EP differentiation. |
Barnstead GenPure xCAD Plus |
Thermo Fisher Scientific |
50136165 |
Water purification system; others can be readily substituted |
Bovine Serum Albumin solution, 7.5% in DPBS, sterile-filtered, BioXtra, suitable for cell culture |
Fisher Scientific |
A8412 |
To preserve cell viability when FACs sorting |
CD34-PE, human (clone: AC136) |
Miltenyi Biotec |
130-098-140 |
Antibody used for FACs isolation of iPSC-EPs |
CHIR99021 |
LC Laboratories |
C-6556 |
Induces the formation of mesoderm from pluripotent stem cells |
Collagen I Rat Tail High Protein 100 mg |
VWR |
354249 |
Main component of the 3D microenvironment |
Conical centrifuge tubes (15/50 mL) |
Fisher Scientific |
14-959-49D/A |
Used to store and mix relatively large volumes of reagents and cell culture media |
DAPI (4’,6-Diamidino-2-Phenylindole, Dihydrochloride) |
Thermo Fisher Scientific |
D1306 |
To counterstain and visualize cell nuclei |
DMEM/F12 |
Thermo Fisher Scientific |
11320–082 |
For dilution of Matrigel and thawing of pluripotent stem cells |
Dulbecco’s phosphate-buffered saline (DPBS) |
ThermoFisher |
14190–250 |
To wash monolayer cultures |
EDTA |
Sigma-Aldrich |
E8008 |
For passaging of pluripotent stem cell colonies and to prevent cell aggregation when FACs sorting |
Endothelial Cell Growth Medium 2 |
PromoCell |
C-22011 |
Promotes endothelial cell viability and proliferation |
Essential 8 Medium |
Thermo Fisher Scientific |
A1517001 |
For maintenance of pluripotent stem cells |
Glycine,BioUltra, for molecular biology, >=99.0% (NT) |
Sigma-Aldrich |
50046 |
Neutralizes remaining detergent |
L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate,>=95% |
Sigma-Aldrich |
A8960 |
Component of iPSC-EP differentiation medium |
MATLAB |
MathWorks |
1.8.0_152 |
Multi-paradigm numerical computing environment (free available at most academic institutions) |
Matrigel Matrix GFR PhenolRF Mouse 10 mL (gelatinous protein mixture) |
Fisher Scientific |
356231 |
Diluted in DMEM/F12 to coat plates for iPSC-EP differentiation |
Medium-199 10X |
Thermo Fisher Scientific |
1825015 |
Used to balance final hydrogel osmolarity and pH |
Microcentrifuge tubes (1.7 mL) |
VWR |
87003–294 |
Stores small volumes of reagents |
Phosphate-buffered saline (PBS) |
Sigma-Aldrich |
P3813 |
The main ingredient of the immunostaining solutions |
Penicillin-Streptomycin (10,000 U/mL) |
Thermo Fisher Scientific |
15140122 |
Antibiotic used after sorting to remove possible contamination from FACS instrument |
Recombinant Human VEGF 165 Protein |
R&D Systems |
293-VE |
Mitogen that stimulates endothelial cell proliferation and tubulogenesis |
Rhodamine phalloidin |
Themo Fisher Scientific |
R415 |
To identify F-actin deposition and therfore outline the borders of the vascular networks |
Triton X-100 (nonionic surfactant) |
Sigma-Aldrich |
X-100 |
Detergent used to gently permeabilize cells |
Tween-20 (emulsifying reagent) |
Fisher Scientific |
BP337 |
Increases the binding specificity of the added antibodies |
VE-Cadherin (F-8) |
Santa Cruz Biotechnology |
sc-9989 |
To identify 3D endothelial lumen in collagen hydrogels |
Vitronectin |
ThermoFisher |
A14700 |
For maintenance of pluripotent stem cells |
Y-27632 |
Selleck Chemicals |
S1049 |
Preserves pluripotent stem cell and iPSC-EP viability when dissociated and re-seeded |