KC-3567

CHOK1-chimeric-DLL3-Cell-Line

46091

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Background of CHOK1-chimeric-DLL3-Cell-Line

DLL3 (Delta-like protein 3), also known as Delta-like protein 3, is a transmembrane protein that plays a role in cell signaling and is a member of the Notch signaling pathway, which is critical for cell differentiation and development. LL3 is involved in intercellular communication and is one of the ligands of the Notch receptor. It interacts with Notch receptors on adjacent cells to trigger a signaling cascade that can influence cell fate decisions, including cell differentiation and proliferation.

Specifications

Catalog Number	KC-3567
Cell Line Name	CHOK1-chimeric-DLL3-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous DLL3 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% 1640 + 20% FBS + 10% DMSO
Propagation Medium	1640 + 10% FBS + 10 μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:4~1:8 every 2~3 days; seed out at about 1-2 x 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

CHOK1-chimeric-DLL3 cell line was generated using a lentiviral vector expressing the chimeric-DLL3 sequence.

Characterization

Figure 1: Characterization of DLL3 overexpression in CHOK1 stable clones using FACS.

Figure 2: Characterization of chimeric-DLL3 overexpression in the CHO-K1 chimeric-DLL3 stable clone using PCR sequence.

Cell Resuscitation

1.Prewarm the culture medium (1640 supplemented with 10% FBS and 10 μg/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 a 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 9.0 mL complete culture medium.
5.Spin at ~ 125 x g for 5~7 minutes at room temperature and discard the supernatant without disturbing the pellet.
6.Resuspend the cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7.Incubate the flask in an incubator at 37°C, 5% CO2.
8.Split the saturated culture at a ratio of 1:4 ~ 1:8 every 2~3 days; seed out at about 1-2 x 105 cells/ml.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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 x g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3 x106 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

1.1.Owen DH, Giffin MJ, Bailis JM, Smit MD, Carbone DP, He K. DLL3: an emergingtarget in small cell lung cancer. J Hematol Oncol. 2019 Jun 18;12(1):61. doi:10.1186/s13045-019-0745-2. PMID: 31215500; PMCID: PMC6582566.
2.2.Yao J, Bergsland E, Aggarwal R, Aparicio A, Beltran H, Crabtree JS, Hann CL,Ibrahim T, Byers LA, Sasano H, Umejiego J, Pavel M. DLL3 as an Emerging Targetfor the Treatment of Neuroendocrine Neoplasms. Oncologist. 2022 Nov3;27(11):940-951. doi: 10.1093/oncolo/oyac161. PMID: 35983951; PMCID:PMC9632312.
3.3.Rojo F, Corassa M, Mavroudis D, Öz AB, Biesma B, Brcic L, Pauwels P, SailerV, Gosney J, Miljkovic D, Hader C, Wu M, Almarez T, Penault-Llorca F.International real-world study of DLL3 expression in patients with small celllung cancer. Lung Cancer. 2020 Sep;147:237-243. doi:10.1016/j.lungcan.2020.07.026. Epub 2020 Jul 27. PMID: 32745892.