KC-3761

293T-ALPP Cell Line

34730

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Background of 293T-ALPP Cell Line

ALPP (Alkaline Phosphatase, Placental), also known as PLAP, is a well-characterized oncofetal antigen. It is normally expressed in the placenta during pregnancy but is significantly silenced in most healthy adult tissues. Its pathophysiological importance arises from its frequent re-expression in various malignancies, most notably in seminomas and non-seminomatous germ cell tumors, as well as in specific subtypes of ovarian, cervical, and lung cancers. This tumor-restricted expression profile has established ALPP as a classic serum biomarker for diagnosing and monitoring germ cell tumors. Furthermore, its prominent cell-surface localization makes it a compelling and actively investigated target for antibody-based therapeutic modalities, including antibody-drug conjugates (ADCs) and CAR-T cell strategies, aiming to selectively target cancer cells.

Specifications

Catalog Number	KC-3761
Cell Line Name	293T-ALPP Cell Line
Clone Number	3#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous ALPP gene
Quantity	One vial 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 +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T-ALPP cell line was generated using lentivirus expressing ALPP sequence.

Characterization

Figure 1. Characterization of ALPP over-expression in the 293T-ALPP stable clone using qPCR.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 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 1:4-1:8 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

Chakraborty R, Das SR, Roy M, Mukherjee BN, Das SK. The effect of parity on placental weight and birth weight: interaction with placental alkaline phosphatase polymorphism. Ann Hum Biol. 1975 Jul;2(3):227-34. doi: 10.1080/03014467500000801. PMID: 16431676.
Borosky GL. Catalytic activity of human placental alkaline phosphatase (PLAP): insights from a computational study. J Phys Chem B. 2014 Dec 11;118(49):14302-13. doi: 10.1021/jp511221c. Epub 2014 Nov 26. PMID: 25409280.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.