| Field | Specification |
|---|---|
| Mfr No | |
| Species |
The LM/TK- (LMTK-) cell line is derived from murine fibroblasts and is characterized by the lack of thymidine kinase (TK) activity. This cell line is particularly useful in genetic and molecular biology research, where it serves as a model system for studying gene function, DNA replication, and recombination. The absence of TK in these cells allows for the selection of mutants or recombinant cells that have regained TK activity, making them valuable in studies involving TK-deficient mutants and for the selection of TK-positive clones following transfection with exogenous DNA.
This cell line, derived from a sub-line of the L-M mouse fibroblast cell line which is resistant to BUdR, is potentially used for genetic and biochemical studies such as gene transfer and somatic cell hybridization. LM/TK- cells are commonly employed in research involving the herpes simplex virus (HSV) thymidine kinase gene, as they provide a crucial background for the selection of HSV-TK gene transformants. This has significant implications in gene therapy research, where HSV-TK is used in suicide gene therapy strategies to selectively kill cancer cells. Furthermore, these cells are utilized in the production of recombinant viruses and in the analysis of viral gene expression and replication. The LMTK- cell line thus plays a critical role in advancing our understanding of genetic manipulation and the development of therapeutic strategies.
- Antigen expression: H-2k
- Tumorigenic: Yes, in nude mice (Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 1×107 cells).
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 2 times per week
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
