ER-Hoxb8 cells WASP+/+ with cell surface antigens T.Y-1.1

Tool Details

*FOR RESEARCH USE ONLY (for other uses, please contact the licensing team)

• Name: ER-Hoxb8 cells WASP+/+ with cell surface antigens T.Y-1.1
• Alternate name: WASP deficient haematopoietic precursors with cell surface antigens TY-1.1
• Cancer: Sarcoma
• Research fields: Cancer
• Tool sub type: Continuous
• Organism: Mouse
• Tissue: Bone Marrow
• Disease: Blood disorders
• Morphology: Variable amoeboid, elongated spindle-like, or round
• Growth properties: Mixed - adherent and suspension
• Model: Knock-Out
• Model description: WASP deficient haematopoietic precursors using CRISPR/Cas9
• Crispr: Yes
• Conditional: Yes
• Conditional description: Conditional HoxB8 expression under ER promoter enabling transcriptional activation to produce immortalized factor-dependant progenitors
• Description: Immortalised ER-Hoxb8 bone marrow progenitor cells expressing cell surface antigens TY-1.1 used to study macrophage functions with implications in cancer immunotherapies targeting the motility of tumour-associated macrophages
• Application: Scanning electron microscopy, FACS, Fluorescence, Immunofluorescence
• Production details: To generate the ER-Hoxb8 Wasp?/? cells, 106 ER-Hoxb8 progenitor cells were seeded on fibronectin coated six-well culture plates in myeloid medium and transduced with 2?ml of viral suspension (as described above) by spinoculation (1000?g, 90?min, 22°C) in the presence of polybrene (8??g/ml). Polybrene was serially diluted by renewing half of the medium over several days to obtain a final concentration of 1.4??g/ml. Antibiotic selection of transduced ER-Hoxb8 cells was performed 3?days post-inf...
• Biosafety level: 2
• Recommended controls: ER-Hoxb8 cells WASP+/+ without cell surface antigens T.Y-1.1

Applications

• Application: Scanning electron microscopy, FACS, Fluorescence, Immunofluorescence

Handling

• Growth medium: Progenitor Medium: RPMI w/ Glutamax (Gibco 61870044), 10% Heat Inactivated FBS, 1x Antibiotic / Antimycotic (Gibco 15240062), 20 ng/mL GM-CSF (Miltenyi 130-095-735), 0.5 ?M ?-Estradiol (Sigma-Aldrich E2758-250MG).
• Temperature: 37° C
• Atmosphere: 5% CO2
• Shipping conditions: Dry ice
• Storage conditions: Liquid Nitrogen
• Subculture routine: If medium is exhausted, spin at RT for 5 min at 300g. Discard medium and resuspend cells in fresh medium. On Mondays and Wednesdays: seed 1.105 cells/mL in new progenitor medium. On Fridays: seed 5.104 cells/mL in new progenitor medium.
• Cultured in antibiotics: Gibco™ Antibiotic-Antimycotic

Related Tools

• Related tools: ER-Hoxb8 cells WASP+/+ without cell surface antigens T.Y-1.1

References

• Accarias et al. 2020. J Cell Sci, 133 (5): jcs236703, PMID: 31964707