| Field | Specification |
|---|---|
| Organism | |
| Applications | |
| Tissue | |
| Growth properties | |
| Biosafety level | |
| Product format | Frozen |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Overview
This cell line is an immortalized neutrophil progenitor cell line dervied from C57Bl/6 Cas9-GFP transgenic mice. Cells express GFP at low concentrations that are detectable by flow cytometry (note: it can be impossible to see GFP expression by fluorescent microscopy).
Key elements and design rationale
- Model identity: Immortalized Neutrophil Cells (mSB8e-C57-Cas9GFP) is supplied as an engineered cell line derived from Mouse bone marrow.
- Growth properties: Suspension, round
- Growth conditions: PriCoat™ T25 Flasks (G299) are recommended for optimal cell culture; passage to new plastic with fresh media at least every 4 days. The base medium for this cell line is PriGrow II (TM002) supplemented with 10% FBS + 100 ng/ml Stem Cell Factor (SCF) (Z200595) + 0.5 μM ß-estradiol + 1% Penicillin/Streptomycin (G255),37.0°C, 5% CO₂. ß-estradiol is required for cell maintenance but must be completely removed for cell differentiation. Note: cells are sensitive to freeze thaw conditions. Do not thaw cells completely; immediately spin down cells and re-suspend in complete growth medium supplemented with 15% FBS to help cells recover for the first 24 hours. For information regarding cell differentiation, please refer to the Differentiation Protocol PDF under the Documents Tab.
- Engineering / immortalization: Gfp reporter expression.
- Product format: Frozen, BSL-2
This cell-based model is generally used in immunology, hematology, and signaling studies.
Biological background
GB8e-C57-Cas9GFP cells are transduced with ER-HoxB8. Cells are capable of differentiation into mature neutrophils. Additional cell lines within this panel include: Cat. T0200 - Immortalized Myeloid Granulocyte Cell Line (ECoM-G) Cat. T0201 - Immortalized Myeloid Monocyte Cell Line (ECoM-M) Cat. T0202 - Immortalized Myeloid Neutrophil Cells (mSB8e-C57STEM-ERKO) Cat. T0203 - Immortalized Myeloid Monocyte Cells (mGB8e-C57STEM-ERKO) Cat. T0204 - Immortalized Neutrophil Cells (mSB8e-C57-Cas9GFP) Cat. T0205 - Immortalized Myeloid Monocyte Cells (mGB8e-C57-Cas9GFP) Cat. T0207 - Immortalized Wild-type Monocyte Progenitor Cells (mGB8e-C57) Cat. T0209 - Immortalized Wild-type Neutrophil Progenitor Cells (mSB8e-C57)
Research relevance and current trends
- Engineered cell lines are widely used for reporter-based readouts, perturbation studies, and assay optimization in reproducible culture systems.
- Reporter or transgene-bearing models are often compared with matched parental or control cells to interpret signal changes in context.
- Expression trends are typically evaluated alongside passage number, selection pressure, and baseline growth behavior.
Common research applications
- Routine expansion and maintenance of a defined cell model for downstream in vitro experiments.
- Phenotype, signaling, or marker-expression studies performed under standardized culture conditions.
- Cell-based assay development in which passage number, growth surface, and medium composition are tracked as experimental variables.
Changes in morphology, growth rate, viability, or reporter signal are typically interpreted together with passage history, culture matrix, and the specified growth conditions for the model.
Notes for experimental interpretation
- Morphology, doubling behavior, and reporter or marker output can shift with passage number, substrate choice, and medium composition; these variables should be recorded alongside experimental readouts.
- Matched controls such as parental cells, untreated cultures, or parallel cultures maintained under identical conditions help distinguish background effects from biology of interest.
Culture and product details
- Growth Conditions: PriCoat™ T25 Flasks (G299) are recommended for optimal cell culture; passage to new plastic with fresh media at least every 4 days. The base medium for this cell line is PriGrow II (TM002) supplemented with 10% FBS + 100 ng/ml Stem Cell Factor (SCF) (Z200595) + 0.5 μM ß-estradiol + 1% Penicillin/Streptomycin (G255),37.0°C, 5% CO₂. ß-estradiol is required for cell maintenance but must be completely removed for cell differentiation. Note: cells are sensitive to freeze thaw conditions. Do not thaw cells completely; immediately spin down cells and re-suspend in complete growth medium supplemented with 15% FBS to help cells recover for the first 24 hours. For information regarding cell differentiation, please refer to the Differentiation Protocol PDF under the Documents Tab.
- Split Ratio: 1:10-1:20 every 3-4 days
- Population Doubling Time (h): 24
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🧊 Thawing Protocol
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
- Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 500xg for 5 minutes.
- Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
- Incubate the cells at the recommended conditions. Note: The cells will experience significant death within 24 hours after thawing, with only a small number surviving—this is an inherent characteristic of the cell line. However, once the viable cells recover, they proliferate rapidly. Recommend to perform hemi-depletions of the media to aid in recovery during the initial period. Media and culture vessels must be changed every 2-4 days.
🔬 Subculture Protocol
- Simply add fresh complete media directly to the culture. Do not allow cell density to exceed 1x10⁶ cells/ml.
- Alternatively, replace complete growth media by centrifuging cells and re-suspending the cell pellet in fresh complete media, and add appropriate aliquots of the cell suspension to new culture vessels, as desired.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
Please refer to our Cell Handling and Thawing Guidelines for detailed instructions on receiving, thawing, and culturing live cells:
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
Why are these cells classified as biosafety level II?
We follow the CDC-NIH recommendations that all mammalian sourced products should be handled at the Biological Safety Level 2 to minimize exposure of potentially infectious products. This information can be found in 'Biosafety in Microbiological and Biomedical Laboratories' (1999). Your institution's Safety Officer or Technical Services will be able to make the call as to whether BioSafety Level I is possible with these cells at your site, if desired.
What is your warranty or return policy?
Our warranty and return policy is outlined in abm’s Terms and Conditions, including details on product quality, limitations, and claims.
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
The number of times cells can divide depends on the cell type:
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Please refer to the Growth Conditions section of your specific product page. This section will indicate whether Applied Cell Extracellular Matrix (G422), PriCoat™ flasks, or both are recommended for optimal cell attachment and growth, as requirements may vary by cell type.
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
Sykes, D. B., & Kamps, M. P. (2003). Estrogen-regulated conditional oncoproteins: tools to address open questions in normal myeloid cell function, normal myeloid differentiation, and the genetic basis of differentiation arrest in myeloid leukemia. Leukemia & lymphoma, 44(7), 1131–1139. https://doi.org/10.1080/1042819031000063444
Wang, G. G., Calvo, K. R., Pasillas, M. P., Sykes, D. B., Häcker, H., & Kamps, M. P. (2006). Quantitative production of macrophages or neutrophils ex vivo using conditional Hoxb8. Nature methods, 3(4), 287–293. https://doi.org/10.1038/nmeth865
Groß, C. J., Mishra, R., Schneider, K. S., Médard, G., Wettmarshausen, J., Dittlein, D. C., Shi, H., Gorka, O., Koenig, P. A., Fromm, S., Magnani, G., Ćiković, T., Hartjes, L., Smollich, J., Robertson, A. A. B., Cooper, M. A., Schmidt-Supprian, M., Schuster, M., Schroder, K., Broz, P., … Groß, O. (2016). K+ Efflux-Independent NLRP3 Inflammasome Activation by Small Molecules Targeting Mitochondria. Immunity, 45(4), 761–773. https://doi.org/10.1016/j.immuni.2016.08.010
Lail, S. S., Arnold, C. R., de Almeida, L. G. N., McKenna, N., Chiriboga, J. A., Dufour, A., Warren, A. L., & Yates, R. M. (2022). Hox-driven conditional immortalization of myeloid and lymphoid progenitors: Uses, advantages, and future potential. Traffic (Copenhagen, Denmark), 23(11), 538–553. https://doi.org/10.1111/tra.12869
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