KC-3725

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

33902

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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	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	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.

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.