KC-3929

293T-KLK3-Cell-Line

40326

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Background of 293T-KLK3-Cell-Line

KLK3, also known as prostate-specific antigen (PSA).Prostate-specific antigen (PSA), a widely used biomarker for prostate cancer (PCa), is encoded by a kallikrein gene (KLK3, kallikrein-related peptidase 3).Prostate cancer shows remarkable clinical heterogeneity, which manifests in spatial and clonal genomic diversity.The prostate produces several proteases, the most abundant ones being kallikrein-related peptidase 3 (KLK3, PSA) and KLK2 (hK2), which are potential targets for tumor imaging and treatment.

Specifications

Catalog Number	KC-3929
Cell Line Name	293T-KLK3-Cell-Line
NCBI/UniProt Accession Number	NM_001648.2
Clone Number	4#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous KLK3 gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1 μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T-KLK3 Cell Line was generated using a lentiviral vector expressing the KLK3 sequence.

Characterization

Figure 1: Characterization of KLK3 overexpression in the 293T-KLK3 stable clone using qPCR.

Figure 2: Characterization of KLK3 overexpression in the 293T-KLK3 stable clone using DNA sequencing.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL Puromycin)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Koistinen H, Künnapuu J, Jeltsch M. KLK3 in the Regulation of Angiogenesis-Tumorigenic or Not? Int J Mol Sci. 2021 Dec 17;22(24):13545.
Chen S, Zhu G, Yang Y, Wang F, Xiao YT, Zhang N, Bian X, Zhu Y, Yu Y, Liu F, Dong K, Mariscal J, Liu Y, Soares F, Loo Yau H, Zhang B, Chen W, Wang C, Chen D, Guo Q, Yi Z, Liu M, Fraser M, De Carvalho DD, Boutros PC, Di Vizio D, Jiang Z, van der Kwast T, Berlin A, Wu S, Wang J, He HH, Ren S. Single-cell analysis reveals transcriptomic remodellings in distinct cell types that contribute to human prostate cancer progression. Nat Cell Biol. 2021 Jan;23(1):87-98.
Koistinen H, Närvänen A, Pakkala M, Hekim C, Mattsson JM, Zhu L, Laakkonen P, Stenman UH. Development of peptides specifically modulating the activity of KLK2 and KLK3. Biol Chem. 2008 Jun;389(6):633-42.
Rodriguez S, Al-Ghamdi OA, Burrows K, Guthrie PA, Lane JA, Davis M, Marsden G, Alharbi KK, Cox A, Hamdy FC, Neal DE, Donovan JL, Day IN. Very low PSA concentrations and deletions of the KLK3 gene. Clin Chem. 2013 Jan;59(1):234-44.