MOLT-4

MOLT-4 is a T lymphoblast cell line derived from the peripheral blood of a 19-year-old male patient with Acute lymphoblastic leukemia (ALL) in relapse in 1971. It is a sister cell line of MOLT-3, while MOLT-4 shows an unusual T-cell antigen receptor gamma-chain gene (T-gamma) rearrangement. MOLT-4 cells have a doubling time of around 30 hours, grow in suspension, and are tumorigenic in untreated nude mice, anti-lymphocyte serum-treated mice, and x-irradiated mice.

MOLT-4 cells have a hypertetraploid chromosome number with the modal chromosome number of 95 occurring in 24% of cells but show stable and recurrent structural abnormalities of chromosomes and longer telomere length. MOLT-4 expresses a variety of T cell markers including CD1, CD2, CD3A, CD3B, CD3C, CD4, CD5, CD6, and CD7. They also express high levels of terminal deoxynucleotidyl transferase (TdT).

The MOLT-4 cell line does not produce immunoglobulin or Epstein-Barr virus. The patient from whom the cells were derived had received prior multidrug chemotherapy. There is a G -> A mutation at codon 248 of the p53 gene, and P53 is not expressed. The line was initially contaminated with mycoplasma but has since been cured with antibiotics.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.