KC-3846

MIAPaCa2-Luc2 Cell Line

34894

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Background of MIAPaCa2-Luc2 Cell Line

Luciferase, a bioluminescent oxidoreductase enzyme, catalyzes photon emission (400-620nm) through oxidation of its luciferin substrate. Its ultra-high sensitivity (detecting gene expression at single-cell resolution) and zero endogenous background establish it as a core reporter tool in oncology research. The broad dynamic range (linear correlation between signal intensity and cell number/gene activity) supports studies on weak promoters and microRNA regulation, while superior in vivo penetration (red-shifted mutants >600nm wavelength reach 3-4cm depth) enables non-invasive real-time monitoring of tumor metastasis, drug response, and PROTAC degradation kinetics. Dual-reporter systems (Firefly + Renilla) ensure precise dissection of cancer pathway mechanisms via internal normalization, and bioluminescence imaging (BLI) quantifies therapeutic efficacy in xenograft models.

Specifications

Catalog Number	KC-3846
Cell Line Name	MIAPaCa2-Luc2 Cell Line
Clone Number	2#
Host Cell Line	MIAPaCa2
Description	Stable MIAPaCa2-Luc2 cell line expressing exogenous Luc2 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 300μg/mL HygromycinB
Selection Marker	HygromycinB
Morphology	attached epithelial with floating rounded cells
Subculture	Split saturated culture 1:2-1:3 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 40 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

MIAPaCa2-Luc2 Cell Line was generated using a lentiviral vector expressing the Luc2 sequence.

Characterization

Figure 1: Characterization of luciferase expression in MIAPaCa2 using the Bright-Lite Luciferase Assay System.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 300μg/mL HygromycinB)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 1:2-1:3 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Greer LF, Szalay AA (2002). Imaging of light emission from the expression of luciferases in living cells and organisms: a review. Luminescence. 17 (1): 43–74. doi:10.1002/bio.676. PMID 11816060.