| Field | Specification |
|---|---|
| Product Format | Frozen |
| Product Type | |
| Shipping | |
| Storage |
Overview
The Immortalized Human T Cells are genetically modified to proliferate indefinitely while retaining their functional characteristics, making them an invaluable tool for immunological research and therapeutic development. These cells offer a consistent and reliable model for studying T cell biology, including mechanisms of activation, signaling pathways, and immune responses.
Key elements and design rationale
- Model identity: Immortalized Human T Cells is supplied as an immortalized cell line derived from Human blood.
- Growth properties: Suspension, round; cells grow as round clumps
- Growth conditions: For optimal initiation of healthy cell cultures, we recommend using a 24-well plate. Once established, cells can be expanded in PriCoat™ T25 Flasks (G299) for continued growth. PrimGrow T Xpan™ Medium Kit (TM182) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. T Xpan™ Cell Activator (CD3/CD28/CD2) (TM139) is required for T cell activation upon thawing. Note 1: refer to the detailed "Thawing Protocol" and "Subculturing Protocol" sections of the Datasheet for complete instructions regarding handling cells. Do not attempt to work with cells until these sections have been reviewed and understood. Note 2: The post-thaw viability of cells can be low, however, correct handling, as per the protocols below, will aid in cell recovery. Cells should form clumps once activated. We recommend thawing frozen cells in 24-well plate and activate before transferring to other culture vessels. Always supplement with fresh reagents. Note 3: We recommend cryopreserving early-passage activated cells as a backup stock for your experiments.
- Engineering / immortalization: Transduction with Lenti-hTERT (LV616) lentivirus and Lenti-Rb siRNA (LV621) lentivirus.
- Product format: Frozen, BSL-2
This cell-based model is generally used in immunology, hematology, and signaling studies. Donor/background information is available for contextual interpretation.
Biological background
Additionally, they are widely used in preclinical studies for developing and testing novel immunotherapies, including CAR-T cell therapies and vaccines. Their ability to be expanded in vitro without senescence allows for large-scale production and in-depth functional assays, facilitating advancements in personalized medicine and immunotherapy research. Donor/background information provided for this product: Healthy donor, no other details disclosed.
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: For optimal initiation of healthy cell cultures, we recommend using a 24-well plate. Once established, cells can be expanded in PriCoat™ T25 Flasks (G299) for continued growth. PrimGrow T Xpan™ Medium Kit (TM182) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. T Xpan™ Cell Activator (CD3/CD28/CD2) (TM139) is required for T cell activation upon thawing. Note 1: refer to the detailed "Thawing Protocol" and "Subculturing Protocol" sections of the Datasheet for complete instructions regarding handling cells. Do not attempt to work with cells until these sections have been reviewed and understood. Note 2: The post-thaw viability of cells can be low, however, correct handling, as per the protocols below, will aid in cell recovery. Cells should form clumps once activated. We recommend thawing frozen cells in 24-well plate and activate before transferring to other culture vessels. Always supplement with fresh reagents. Note 3: We recommend cryopreserving early-passage activated cells as a backup stock for your experiments.
- Immortalization Method: Transduction with Lenti-hTERT (LV616) lentivirus and Lenti-Rb siRNA (LV621) lentivirus.
- Thaw the cryovial rapidly in a 37 °C water bath with gentle agitation for no longer than 2 minutes. Keep the cap above the water level to minimize contamination risk.
- Decontaminate the vial by spraying and wiping the exterior with 70% ethanol. From this step onward, perform all procedures inside a certified biological safety cabinet under strict aseptic conditions.
- Transfer the cell suspension into a 15 ml sterile conical tube containing 5 ml of pre-warmed PrimGrow T Xpan™ Medium (TM182), supplemented at the time of use with 10% FBS and ROCK Inhibitor Y-27632 (TM131). Centrifuge at 125 × g for 5 minutes.Supplementing TM182-B with FBS and Y-27632 during this step improves post-thaw cell viability.
- Carefully aspirate the supernatant without disturbing the cell pellet. Resuspend and plate 1 × 10⁶ cells per well into a 24-well plate in 1 ml of non-supplemented PrimGrow T Xpan™ Medium (TM182). Add 2 µl of T Xpan™ Cell Activator (CD3/CD28/CD2) (TM139) to each well and incubate for 24 hours.
- After 24 hours, observe the culture for cluster formation, which indicates successful activation. If clusters are not yet visible, continue incubation and re-check at 48 hours. Once clusters are observed, directly dilute and transfer the cells into a 6-well plate, adding fresh supplements to support continued expansion.
- Subculture the cells at a 1:2 split ratio once the cell density exceeds 3 × 10⁶ cells/ml. Gradually expand the culture into T25 or T75 flasks as needed for downstream applications.
- To maintain robust proliferation, re-stimulate the culture with fresh antibody-based activator every 7–14 days, using 2 µl of T Xpan™ Cell Activator (TM139)
- Simply add fresh complete media directly to the culture flask. or,
- Remove half of the cell suspension and transfer to a new culture vessel; add additional fresh complete media to both culture vessels.
- Incubate the cells at the recommended conditions.
- Continue to monitor cell growth (cell clumps should continue to grow and become visible in the culture flask). Add fresh complete growth media every 3 days.
- Rapid proliferation typically continues for up to 14 days, during which cells should be passaged every 1–2 days. Afterward, they return to a more stable growth phase.
- If additional expansion is needed after day 14, a second activation (repeat the 24-well setup) may be performed only when cells are healthy. However, avoid repeated stimulations — more than 3–5 rounds can induce T cell exhaustion. Note: Do not centrifuge cells during subculture procedures.
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.
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