| Field | Specification |
|---|---|
| Mfr No | |
| Species |
HEL-299 is a human lung fibroblast cell line derived from an adult individual. This cell line is particularly noted for its finite capacity to propagate in culture, typically entering senescence after approximately ten passages. This characteristic makes HEL-299 a useful model for studying cellular aging and senescence, as well as the dynamics of cell growth and replication under controlled conditions.
In addition to its applications in aging research, HEL-299 also serves as a model for studying signal transduction pathways. Specifically, it has been observed that the expression of the M2 muscarinic receptor in these cells is downregulated following stimulation with protein kinase C. This response highlights the cell line's utility in pharmacological research and in the investigation of mechanisms underlying receptor-mediated signaling and regulation. The alteration in receptor expression following kinase activity can provide insights into cellular responses to external stimuli, potentially aiding in the development of therapeutic strategies targeting similar pathways in various diseases.
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- Receptors expressed: m2 muscarinic receptor
- Protein expression: p53 negative
- Isoenzymes: G6PD, A
- Virus susceptibility: Vesicular stomatitis (Indiana), poliovirus 1
- Reverse transcriptase: Negative
- Karyotype: Normal human male, diploid, stable
- cultureMedium: Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a)
- supplements: Supplement the medium with 10% FBS, 1 ng/mL bFGF
- 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.
- seedingDensity: 1 x 104 cells/cm2
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- 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.
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