| Field | Specification |
|---|---|
| Product Format | Frozen |
| Product Type | |
| Shipping | |
| Storage |
Overview
MRC-5 is a diploid human cell line composed of normal lung fibroblasts derived from fetal tissue. These cells are widely used as a model for normal human fibroblasts in virology, cytogenetics, vaccine production, and cellular senescence research. The product is supplied as frozen cells.
Key elements and design rationale
- Biological source: Human (H. sapiens)
- Tissue origin: Lung
- Growth properties: Adherent, fibroblast-like
- Format: Frozen
- Donor history: Male, 14 weeks gestation (fetus), Normal lung tissue
- Biosafety level: II
- Culture context: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. DMEM, High Glucose Medium (TM500) + 10% FBS (*Regular) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.*Do not heat-inactivate
Primary cells provide a biologically relevant starting point for in vitro studies because they retain tissue-linked morphology, growth behavior, and donor-associated characteristics that are often reduced in immortalized systems.
Research relevance and current trends
- Primary-cell models continue to be used when donor context, tissue specificity, and physiologic responsiveness are important.
- Researchers often combine primary cells with co-culture, matrix, or conditioned-media approaches to better reflect native microenvironments.
- Phenotypic characterization and careful tracking of culture conditions remain important for reproducible interpretation.
Common research applications
- Use in tissue-relevant in vitro assays where morphology and donor context matter.
- Measure phenotype, marker expression, or response to defined perturbations over time.
- Incorporate into co-culture or screening workflows requiring primary-cell context.
Product-specific data supplied for this listing
- Growth Conditions: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. DMEM, High Glucose Medium (TM500) + 10% FBS (*Regular) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.*Do not heat-inactivate
- Split Ratio: 1:2-1:3
Notes for experimental interpretation
- Primary cells can show donor-dependent and passage-dependent variation, so morphology, growth rate, and marker expression should be interpreted in the context of the specific lot and culture history.
- Attachment matrix, medium formulation, and gas conditions can materially influence phenotype maintenance and experimental readouts; use the recommended culture system where possible.
- Cryopreserved recovery conditions matter for viability and downstream behavior. We recommend using serum-free CryoGuard™ Freezing Media (TM078) or, if serum is preferred, Cryopreservation Medium (TM024).
- 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.
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.
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