REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Anti-human CD34 FITC (4H11) | eBioscience | Cat# 11-0349-42; RRID: AB_1518732 |
Biological samples | ||
Fresh human umbilical cord blood | StemCyte | https://www.stemcyte.com/ |
Chemicals, peptides, and recombinant proteins | ||
Puromycin Dihydrochloride | Sigma-Aldrich | Cat# P8833; CAS: 58-58-2 |
Bovine Serum Albumin solution | Sigma-Aldrich | Cat# A7034; CAS: 9048-46-8 |
Recombinant Human IL-6 | PeproTech | Cat# 200-06 |
Recombinant Human IL-3 | PeproTech | Cat# 200-03 |
Recombinant Human SCF | PeproTech | Cat# 300-07 |
L-Glutamine (200 mM) | Thermo Fisher Scientific | Cat# 25030-081 |
Sodium Pyruvate (100 mM) | Thermo Fisher Scientific | Cat# 11360-070 |
Penicillin Streptomycin | Thermo Fisher Scientific | Cat# 15-140-122 |
Plasmocin prophylactic | InvivoGen | Cat# ant-mpp |
M-PER Mammalian Protein Extraction Reagent | Thermo Fisher Scientific | Cat# 78501 |
Ficoll Paque Plus | GE Healthcare | Cat# 17-1440-02 |
DMEM, high glucose, with pyruvate | Life Technologies | Cat# 11995-073 |
X-tremeGENE HP DNA Transfection Reagent | Sigma-Aldrich | Cat# 6366546001 |
RetroNectin® Recombinant Human Fibronectin Fragment | Takara Bio Inc. | Cat# T100B |
Trypsin-EDTA 0.05% | Fisher Scientific | Cat# 25-300-120 |
Fetal bovine serum (FBS) | Gemini Bio-Products | Cat# 100-106 |
Dimethyl Sulfoxide (DMSO) | Fisher Bioreagents | Cat# BP231-100; CAS: 67-68-5 |
HEPES (1M) | Thermo Fisher Scientific | Cat# 15630080 |
Polybrene | Sigma-Aldrich | Cat# H9268; CAS: 28728-55-4 |
StemSpan SFEM medium | STEMCELL Technologies | Cat# 09650 |
UltraPure 0.5M EDTA, pH 8.0 | Invitrogen | Cat# 15575020 |
Cell-Tak Cell and Tissue Adhesive | Corning | Cat# 354240 |
Seahorse XF RPMI Medium, pH 7.4 | Agilent Technologies | Cat# 103576-100 |
Glucose Solution | Gibco | Cat# A2494001 |
Sodium bicarbonate | Sigma-Aldrich | Cat# S5761-500G; CAS: 144-55-8 |
Critical commercial assays | ||
CD34 MicroBead Kit, human | Miltenyi Biotec | Cat# 130-046-702 |
Seahorse XF Glycolytic Rate Assay Kit | Agilent Technologies | Cat# 103344-100 |
Bio-Rad Protein (Bradford) Assay Dye Reagent | Bio-Rad | Cat# 5000006 |
Seahorse XFe96 FluxPak mini | Agilent Technologies | Cat# 102601-100 |
Experimental models: Cell lines | ||
293T | ATCC | (CRL-3216); RRID:CVCL_0063 |
Recombinant DNA | ||
pMD2.G | A gift from Dr. Didier Trono | Addgene plasmid # 12259; RRID: Addgene_12259 |
pMDLg/pRRE | (Dull et al., 1998) | Addgene plasmid # 12251; RRID: Addgene_12251 |
pRSV-Rev | (Dull et al., 1998) | Addgene plasmid # 12253; RRID: Addgene_12253 |
pLKO.1-shNS | (Stewart et al., 2003) | Addgene plasmid # 8453 |
pLKO.1-shFTO-1 | Sigma-Aldrich | TRCN0000246247 |
pLKO.1-shFTO-2 | Sigma-Aldrich | TRCN0000246249 |
Software and algorithms | ||
GraphPad Prism | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
Seahorse Wave | Agilent Technologies | https://www.agilent.com/en/product/cell-analysis/real-time-cell-metabolic-analysis/xf-software/seahorse-wave-desktop-software-740897 |
Seahorse XF Glycolytic Rate Assay Report Generators | Agilent Technologies | https://www.agilent.com/en/product/cell-analysis/real-time-cell-metabolic-analysis/xf-software/seahorse-xf-glycolytic-rate-assay-report-generators-740900 |
FlowJo v10 | BD Life Sciences | https://www.flowjo.com/ |
Other | ||
Falcon polystyrene microplates, non-treated | Corning | Cat# 351146 |
Seahorse XFe96 Analyzer | Agilent Technologies | n/a |
Eppendorf Research plus 8-channel adjustable volume pipette, 30–300 μL | Eppendorf | Cat# 3125000052 |
Eppendorf Research plus 8-channel adjustable volume pipette, 10–100 μL | Eppendorf | Cat# 3125000036 |
Matrix Reagent Reservoirs | Thermo Scientific | Cat# 8094 |