KC-6660

LNCaP-ACP3 Cell Line

64239

Home » LNCaP-ACP3 Cell Line

Background of LNCaP-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. Diseases associated with ACP3 include Prostatitis and Clear Cell Adenocarcinoma.

Specifications

Catalog Number	KC-6660
Cell Line Name	LNCaP-ACP3 Cell Line
NCBI/UniProt Accession Number	NM_001134194.1
Clone Number	4#
Host Cell Line	Human LNCaP Cell Line
Description	Stable LNCaP cell line expressing exogenous human ACP3 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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS +2μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:2-1:3 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 56-84 hours
Mycoplasma Status	Negative

Cell Line Generation

LNCaP-ACP3 cell line was generated using a lentiviral vector expressing the human ACP3 sequence.

Characterization

Figure 1: Characterization of endogenous human ACP3 expression in LNCaP-ACP3 cells using FACS.

Figure 2: Characterization of human ACP3 in the LNCaP-ACP3 stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (RPMI1640+10% FBS +2μg/mL Puromycin) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:2-1:3 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Erbas H, Erten O, Irfanoglu ME. Prostatic acid phosphatase in breast cyst fluid. Malays J Pathol. 2007 Dec;29(2):95-9. PMID: 19108401.
2. 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.
3. Arnold F, Schnell J, Zirafi O, Stürzel C, Meier C, Weil T, Ständker L, Forssmann WG, Roan NR, Greene WC, Kirchhoff F, Münch J. Naturally occurring fragments from two distinct regions of the prostatic acid phosphatase form amyloidogenic enhancers of HIV infection. J Virol. 2012 Jan;86(2):1244-9. doi: 10.1128/JVI.06121-11. Epub 2011 Nov 16. PMID: 22090109; PMCID: PMC3255800.

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.