Table 1.
Key resources table
| Brand | Product | Cat No. | Concentration | |
|---|---|---|---|---|
| IMGL generation | ||||
| Medium and reagents | Gibco™ | Essential 8™ Basal Medium (500 mL) | A1516901 | NA |
| Gibco™ | Essential 8™ Supplement (50X) (10 mL) | A1517101 | 1x | |
| Stemcell™ | mTeSR™1 Basal Medium (400 mL) | #85851 | NA | |
| Stemcell™ | mTeSR™1 Supplement (5X) (100 mL) | #85852 | 2.0 mL/mL | |
| Gibco™ | RevitaCell™ Supplement (100X) (5 mL) | A2644501 | 1x | |
| Merck | In-solution Rock Inhibitor (500 μg/296 uL) | Y-27632 | 10 μM | |
| Lonza | XVIVO ™ 15 | BE02-060Q | NA | |
| Thermofisher | 2-mercaptoethanol | 31350010 | 0.055 mM | |
| Gibco™ | GlutaMax | 35050061 | 2 mM 1% | |
| Gibco™ | Penicillin/Streptomycin | 15070063 |
100 mg/mL 100 U/mL |
|
| Gibco™ | DPBS 1x | 14190250 | NA | |
| Sigma-Aldrich | Accutase Solution | A6963 | NA | |
| Sigma-Aldrich | EDTA (Ethylenediaminetetraacetic acid disodium salt solution | E7889-100mL | 0,5 mM | |
| Gibco™ | Advanced DMEM/F12 1x Reduced Serum medium (1:1) 500 mL 3.15 g/L | 12634-010 | NA | |
| Plastic equipment | Greiner Bio-one | 6 Well culture plate | 657 160 | NA |
| Corning |
96-well Ultra Low attachment plate U-bottom |
7007 | NA | |
| STEMCELL | Reversible strainer 37μm large | 27250 | NA | |
| Maturation medium supplements | Sigma | N-acetylcysteine | A0737 | 5 μg/mL |
| Sigma | 1-Thioglycerol | M6145 | 400 μM | |
| Sigma | Heparan sulfate | H7640 | 1 μg/mL | |
| Gibco™ | NEAA | 11140-050 | 1% | |
| Gibco™ | B27 | 17504-044 | 2% | |
| Gibco™ | N2 | 17502-048 | 0.5% | |
| Growth Factors | Peprotech | Recombinant Human BMP-4 50 μg | 125-05 | 50 ng/mL |
| Peprotech | Recombinant Human VEGF165 50 μg | 100-20 | 50 ng/mL | |
| Peprotech | Recombinant Human SCF 50 μg | 300-07 | 20 ng/mL | |
| Peprotech | Recombinant Human M-CSF (CSF1) 50 μg | 300-25 |
50 ng/mL 25 ng/mL |
|
| Peprotech | Recombinant Human IL-3 50 μg | 200-03 | 50 ng/mL | |
| Peprotech | Recombinant Human FLT3-ligand 50 μg | 300-19 | 50 ng/mL | |
| Peprotech | Recombinant HumanTPO 50 μg | 300-18 | 5 ng/mL | |
| Peprotech |
Recombinant Human GM-CSF 50 μg |
300-03 | 25 ng/mL | |
| Peprotech | Recombinant Human Interleukin-34 | 200-34 | 100 ng/mL | |
| Peprotech | Recombinant Human CX3CL1 | 300-31 | 25 ng/mL | |
| Peprotech | Recombinant Human TGF-β1 | 100-21 C | 25 ng/mL | |
| R&D systems | CD200 | 2724-CD-050 | 50 ng/mL | |
| Medium and reagents | ||||
| Gibco™ |
DMEM High Glucose 4.5 g/L |
42430025 | NA | |
| Agilent, | Seahorse XF base medium | 102353-100 | ||
| Hyclone | fetal bovine serum (FBS) | SV30160 | 10% | |
| Thermo Fisher | Sodium Pyruvate | 15070063 | 1 mM | |
| Sigma‐Aldrich | Lipopolysaccharide (LPS) Escherichia coli | L4391 |
1 mg/kg (In vivo) 250 ng/ml (In vitro) |
|
| Lonza | DPBS | BE17512F | ||
| Sigma Aldrich | Cytochalasin D | C8273 | 10uM | |
| Thermo Fischer | pHrodo™ Red E. coli BioParticles™ Conjugate for Phagocytosis | p35361 | 1ug/0.1 ml | |
| Sigma Aldrich | Paraformaldehyde (PFA) | P6148 | 4% | |
| Sigma Aldrich | Tritox X-100 | 11332481001 | 0.1% | |
| Sigma Aldrich | Bovine Serum Albumine (BSA) | A7030 | 5% | |
| Lonza | MycoAlert® PLUS Mycoplasma Detection Kit | LT07-710 | NA | |
| Sigma Aldrich | Fluoroshield™ with DAPI | F6057 | NA | |
| Sigma Aldrich | Glucose | G8270 | 10 mM | |
| Thermo Fischer | Glutamine | 25030024 | 2 mM | |
| Sigma Aldrich | Oligomycin | 04876 | 2.5 uM | |
| Sigma Aldrich | FCCP | C2920 | 0.5 uM | |
| Sigma Aldrich | Rotenone | R8875 | 0.5 uM | |
| Sigma Aldrich | Antimycin A | A8674 | 0.5 uM | |
| Sigma Aldrich | β-Nicotinamide adenine dinucleotide hydrate (NAD) | N7004 | 25 mM | |
| Sigma Aldrich | Lactate dehydrogenase | L2500 | 100 U/mL | |
| BIOKE | NucleoSpin RNA Isolation kit | 740955 | ||
| Plastic equipment | ||||
| Agilent | 96-well E-plate | 5232368001 | ||
| Agilent Technologies | Extracellular Flux Assay Kit | |||
| Greiner bio-one | Cellstrar tubes 15 ml | 188285 | ||
| Antibodies | ||||
| Wako | IBA1, Rabbit Lot: PTR2404 | 019-19741 | (1:500) | |
| Invitrogen | Alexa Fluor 488 Lot: 2256732 | A21206 | (1:2000) | |
| Sigma Aldrich | Fluoroshield™ with DAPI | F6057 | NA | |
| Oligonucleotides | ||||
| Biolegio |
NANOG F: CAATGGTGTGACGCAGAAGG R: TGCACCAGGTCTGAGTGTTC |
|||
| Biolegio |
NeuN F: ACGATCGTAGAGGGACGGAA R: TTTAGCTTCCAGCCGTTGGT |
|||
| Biolegio |
P2RY12 F: TCAGATTACAAGAGCACTCAAGACT R: CTGCAGAGTGGCATCTGGTAT |
|||
| Biolegio |
CD11C F: GGGGTGCTGTCTACCTGTTT R: GGGGTGCTGTCTACCTGTTT |
|||
| Biolegio |
HPRT F: TCATTATGCTGAGGATTTGGAAA R: GGCCTCCCATCTCCTTCATC |
|||
| Biolegio |
TMEM119 F: CAAGGAACTGGTCCTGGGG R: CAGGAGCAGCAACAGAAGGA |
|||
| Biolegio |
HBMS F: TGGACCTGGTTGTTCACTCCTT R: CAACAGCATCATGAGGGTTTC |
|||
Not applicable is shown as NA.