| Field | Specification |
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| Applications | |
| Tissue | |
| Growth properties | |
| Biosafety level | |
| Product format | Frozen |
| Storage | |
| Shipping | |
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Overview
Immortalized Rat Embryonic Fibroblasts (18IM) are derived from rat embryonic fibroblasts isolated from 13 to 14 day old fetuses which were transfected with a plasmid containing S18-2. Transfected cells were selected for with 0.5 mg/mL G418.
Key elements and design rationale
- Model identity: Immortalized Rat Embryonic Fibroblasts (18IM) is supplied as an immortalized cell line derived from Rat embryo.
- Growth properties: Adherent, polygonal
- Growth conditions: Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow V (TM015) + 5% FBS + 0.5 mg/ml G418 (G271) +1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂
- Product format: Frozen, BSL-2
This cell-based model is generally used in vascular biology, signaling, and assay development studies.
Biological background
18IM cells display embryonic stem cell-like properties after immortalization and subsequent loss of mesodermal markers thereafter. Loss of contact inhibition was also observed and cells were also capable of showing anchorage-independent growth. Immunostaining showed the presence of stem cell markers, such as SSEA-1, and the loss of mesodermal markers, such as vimentin and smooth muscle actin. RT-PCR also confirmed the loss of mesodermal markers, FUT4 and vimentin, and an up-regulation of stem cell markers, Sox2 and Oct3/4. This cell line may be useful as a model for the role of S18-2 in other cell types and further studies in the binding partners of S18-2 in differentiated tissues. Western blot analysis for presence of S18-2 in cells.
Research relevance and current trends
- Cultured cell-line models remain central to in vitro studies of phenotype, signaling, and pathway regulation under controlled conditions.
- Researchers commonly compare morphology, growth rate, and marker expression across media formulations, treatments, or time courses.
- Interpretation is generally strengthened by using matched controls, consistent passage handling, and appropriate culture surfaces.
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: Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow V (TM015) + 5% FBS + 0.5 mg/ml G418 (G271) +1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂
- Split Ratio: 1:2 to 1:3
- Seeding Density (cells/cm²): 20,000 – 100,000
- 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 125xg for 5-7 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.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.
- Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
- Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. 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?
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?
What is your warranty or return policy?
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?
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?
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.
