KC-6328

LNcap-ACP3-KO cell line

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Background of LNcap-ACP3-KO cell line

Acid phosphatase 3 (ACP3), a tyrosine phosphatase primarily expressed in immune cells, particularly T cells, negatively regulates the T cell receptor signaling pathway by dephosphorylating key signaling molecules, thereby maintaining immune homeostasis. Research indicates that ACP3 plays a crucial role in preventing excessive T-cell activation and autoimmune responses. Consequently, the ACP3 gene knockout model has become a vital tool for studying autoimmune diseases and cancer immunotherapy. By knocking out the ACP3 gene, its inhibitory effect on T-cell activation can be lifted, potentially enhancing anti-tumor immune responses. Current research focuses on utilizing ACP3 knockout strategies to develop novel adoptive T-cell therapies, aiming to provide new targets and approaches for tumor immunotherapy.

Specifications

Catalog Number	KC-6328
Cell Line Name	LNcap-ACP3-KO cell line
Clone Number	2A3
Host Cell Line	LNcap
Description	Stable LNcap cell clone with human ACP3 gene knockout
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
Selection Marker	NA
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:3-1:4 every 3-4 days; seed out at about 1-3 x 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 36 hours
Mycoplasma Status	Negative

Cell Line Generation

LNcap-ACP3-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of LNcap-ACP3-KO cell line stable clone using PCR sequencing.

Figure 2:Characterization of LNcap-ACP3-KO cell line stable clone using RT-PCR sequencing.

Figure 3:Characterization of LNcap-ACP3-KO cell line stable clone using western blot.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10%FBS)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 1:3-1:4 every 3-4 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

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.
Farmer R, Thomas CM, Winn PJ. Structure, function and dynamics in acyl carrier proteins. PLoS One. 2019 Jul 10;14(7):e0219435. doi: 10.1371/journal.pone.0219435. PMID: 31291335; PMCID: PMC6619796.