CELL LINES
Contributor Information
- Name Véronique Le Cabec, Thibaut Sanchez, Frederic Lagarrigue
- Institute Toulouse Tech Transfer
- Primary citation Accarias et al. 2020. J Cell Sci, 133 (5): jcs236703, PMID: 31964707
Tool Details
- Tool name: ER-Hoxb8 cells WASP+/+ without cell surface antigens T.Y-1.1
- Alternate names: WASP deficient haematopoietic precursors without cell surface antigens TY-1.1
- Tool type: Cell Lines
- Tool sub-type: Continuous
- Organism: Mouse
- Tissue: Bone Marrow
- Cancer type: Sarcoma
- Disease: Haematological diseases
- 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
- Application: Scanning electron microscopy, FACS, Fluorescence, Immunofluorescence
- 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
- Research area: Cancer
- 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-infection with puromycin (10??g/ml) and maintained for at least 2?weeks. Immortalized ER-Hoxb8 Wasp?/? cells were maintained and enriched in myeloid medium with puromycin (2.5??g/ml) by serial passages of non-adherent cells every 34?days into new six-well culture plates. The same protocol was used to generate Wasp+/+ control ER-Hoxb8 cells, except that empty viruses were used.' (Accarias et al. 2020. J Cell Sci, 133 (5): jcs236703, PMID: 31964707)
- For Research Use Only
Target Details
Application Details
- 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
- Storage conditions: Liquid Nitrogen
- Shipping conditions: Dry ice
- Cultured in antibiotics?: Gibco Antibiotic-Antimycotic
- Biosafety level: BSL-2
- 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.
Documentation
Related Tools
References
- • Accarias et al. 2020. J Cell Sci, 133 (5): jcs236703, PMID: 31964707
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