KC-5754

HT1080-ACP3 Cell Line

59019

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Background of HT1080-ACP3 Cell Line

The ACP3 gene encodes a lysosomal acid phosphatase that catalyzes the hydrolysis of orthophosphoric monoesters, a critical reaction in phosphate metabolism. A key regulatory feature of this gene is its control by androgens, leading to its specific secretion by prostate epithelial cells, which implicates its potential role in prostate physiology and related pathologies. The discovery of an alternatively spliced isoform containing a transmembrane domain localizes this variant to the plasma membrane-endosomal-lysosomal pathway. This distinct subcellular localization shifts the research focus from a purely lysosomal enzyme to a potential cell-surface modulator of phosphorylated signaling molecules. Consequently, current investigations center on how this membrane-associated ACP3 isoform may influence phosphoprotein dynamics at the cell surface, thereby affecting signal transduction pathways in hormone-responsive cancers like prostate cancer.

Specifications

Catalog Number	KC-5754
Cell Line Name	HT1080-ACP3 Cell Line
Clone Number	22#
Host Cell Line	HT1080
Description	Stable HT1080 cell line expressing exogenous ACP3 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+1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Fibroblast
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

Cell Line Generation

HT1080-ACP3 Cell Line was generated using a lentiviral vector expressing the ACP3 sequence.

Characterization

Figure 1: Characterization of ACP3 overexpression in the HT1080-ACP3 stable clone using FACS.

Figure 1: Characterization of ACP3 overexpression in the HT1080-ACP3 stable clone using PCR.

Cell Resuscitation

Prewarm culture medium (RPMI1640+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% 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

Erbas H, Erten O, Irfanoglu ME. Prostatic acid phosphatase in breast cyst fluid. Malays J Pathol. 2007 Dec;29(2):95-9. PMID: 19108401.
Olsen JS, DiMaio JT, Doran TM, Brown C, Nilsson BL, Dewhurst S. Seminal plasma accelerates semen-derived enhancer of viral infection (SEVI) fibril formation by the prostatic acid phosphatase (PAP248-286) peptide. J Biol Chem. 2012 Apr 6;287(15):11842-9. doi: 10.1074/jbc.M111.314336. Epub 2012 Feb 21. PMID: 22354963; PMCID: PMC3320932.