KC-5891

CHOK1-cyno-ACP3-Cell-Line

60677

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Background of CHOK1-cyno-ACP3-Cell-Line

ACP3 encodes an enzyme that catalyzes the conversion of orthophosphoric monoester to alcohol and orthophosphate. It is synthesized under androgen regulation and is secreted by the epithelial cells of the prostate gland. ACP3 has important clinical significance in prostate cancer.

Specifications

Catalog Number	KC-5891
Cell Line Name	CHOK1-cyno-ACP3-Cell-Line
NCBI/UniProt Accession Number	XM_005545772.5
Clone Number	7#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous cyno-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% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-cyno-ACP3-cell-line was generated using a lentiviral vector expressing the cyno-ACP3 sequence.

Characterization

Figure 1: Characterization of cyno-ACP3 overexpression in the CHOK1-cyno-ACP3 stable clone using qPCR.

Figure 2: Characterization of cyno-ACP3 in the CHOK1-cyno-ACP3 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1.Cao Z, Pu C, Jiang X, Han G, Shen X, Wang W, Ding W, Huang Z, Huang X, Jia B, Lu VX, Tian L, Wu Z, Xiao L. Novel PAP-targeted CAR-T therapy enhances antitumor efficacy through CoupledCAR approach. J Immunother Cancer. 2025 May 31;13(5):e011238. doi: 10.1136/jitc-2024-011238. PMID: 40449956; PMCID: PMC12161386.
2.Kałuża A, Trzęsicka K, Drzyzga D, Ferens-Sieczkowska M. Aberrant Mannosylated and Highly Fucosylated Glycoepitopes of Prostatic Acid Phosphatase as Potential Ligands for Dendritic-Cell Specific ICAM-Grabbing Nonintegrin (DC-SIGN) in Human Seminal Plasma-A Step towards Explaining Idiopathic Infertility. Biomolecules. 2023 Dec 31;14(1):58. doi: 10.3390/biom14010058. PMID: 38254658; PMCID: PMC10813591.
3.Liu X, Yu L, Xia Z, Li J, Meng W, Min L, Li F, Wang X. Purification, identification and Cryo-EM structure of prostatic acid phosphatase in human semen. Biochem Biophys Res Commun. 2024 Apr 2;702:149652. doi: 10.1016/j.bbrc.2024.149652. Epub 2024 Feb 7. PMID: 38341922.