KC-3725

293T-GPC3-KO-1C3-Cell-Line

33902

Home » 293T-GPC3-KO-1C3-Cell-Line

Background of 293T-GPC3-KO-1C3-Cell-Line

GPC3 (Glypican 3) is a Protein Coding gene. Diseases associated with GPC3 include Simpson-Golabi-Behmel Syndrome, Type 1 and Wilms Tumor 1. Among its related pathways are Chondroitin sulfate/dermatan sulfate metabolism and Defective EXT2 causes exostoses 2. Gene Ontology (GO) annotations related to this gene include heparan sulfate proteoglycan binding and peptidyl-dipeptidase inhibitor activity.

Specifications

Catalog Number	KC-3725
Cell Line Name	293T-GPC3-KO-1C3-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone with human GPC3 gene knockout, No.1C3
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	DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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-GPC3-KO-1C3 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of 293T-GPC3-KO-1C3 cell line stable clone using PCR sequencing.

Figure 2: Characterization of 293T-GPC3-KO-1C3 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of 293T-GPC3-KO-1C3 cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM + 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 1:3-1:6 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

Samson N, Ablasser A. The cGAS-STING pathway and cancer. Nat Cancer. 2022 Dec;3(12):1452-1463. doi: 10.1038/s43018-022-00468-w. Epub 2022 Dec 12. PMID: 36510011.
Xie J, Li Y, Shen X, Goh G, Zhu Y, Cui J, Wang LF, Shi ZL, Zhou P. Dampened STING-Dependent Interferon Activation in Bats. Cell Host Microbe. 2018 Mar 14;23(3):297-301.e4. doi: 10.1016/j.chom.2018.01.006. Epub 2018 Feb 22. PMID: 29478775; PMCID: PMC7104992.
Tretiakova M, Zynger DL, Luan C, Andeen NK, Finn LS, Kocherginsky M, Teh BT, Yang XJ. Glypican 3 overexpression in primary and metastatic Wilms tumors. Virchows Arch. 2015 Jan;466(1):67-76. doi: 10.1007/s00428-014-1669-4. Epub 2014 Nov 4. PMID: 25366870.