ER-Hoxb8 cells WASP+/+ without cell surface antigens T.Y-1.1

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

Contributors

Inventor Institute
Véronique Le Cabec, Thibaut Sanchez, Frederic Lagarrigue Toulouse Tech Transfer
Cat. #: 161707
Tool sub type: Continuous
Cancer types: Sarcoma
Research Fields: Cancer
Organism: Mouse
Tissue: Bone Marrow
Model: Knock-Out
Morphology: Variable amoeboid, elongated spindle-like, or round
Growth properties: Mixed – adherent and suspension
Primary citation: Accarias et al. 2020. J Cell Sci, 133 (5): jcs236703, PMID: 31964707
Alternate name: WASP deficient haematopoietic precursors without cell surface antigens TY-1.1
Product 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
CRISPR: Yes
Conditional: Yes
Conditional description: Conditional HoxB8 expression under ER promoter enabling transcriptional activation to produce immortalized factor-dependant progenitors
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…
Model description: WASP deficient haematopoietic precursors using CRISPR/Cas9
disease: Blood disorders
Storage conditions: Liquid Nitrogen
Shipping conditions: Dry ice
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).
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.
Temperature: 37° C
Atmosphere: 5% CO2
Biosafety level: 2
References:

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

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