KC-3192

CHOK1-mouse-SEZ6-Cell-Line

34202

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Background of CHOK1-mouse-SEZ6-Cell-Line

Acts upstream of or within several processes, including activation of protein kinase C activity; excitatory postsynaptic potential; and nervous system development. Located in several cellular components, including dendrite; neuronal cell body; and perinuclear region of cytoplasm. Is expressed in several structures, including central nervous system; genitourinary system; hemolymphoid system gland; liver; and retina. Orthologous to human SEZ6 (seizure related 6 homolog).

Specifications

Catalog Number	KC-3192
Cell Line Name	CHOK1-mouse-SEZ6-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous mouse SEZ6 gene
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	70% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/ml Puromycin
Selection Marker	Puromycin
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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 mouse SEZ6 Cell Line was generated using a lentiviral vector expressing the mouse SEZ6 sequence.

Characterization

Figure 1: Characterization of mouse SEZ6 overexpression in CHOK1 stable clones using FACS.

Figure2: Characterization of endogenous mouse SEZ6 expression in CHOK1 cells using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/mL Puromycin)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 1:4-1:8 every 2-3 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

Dominko K, Rastija A, Smiljanic K, Mladenovic A, Lešnjaković L, Kanazir S, Milanovic D, Hecimovic S. Amyloid-ß plaque formation and BACE1 accumulation in the brains of a 5xFAD Alzheimer's disease mouse model is associated with altered distribution and not proteolysis of BACE1 substrates Sez6 and Sez6L. Mech Ageing Dev. 2022 Oct;207:111726. doi: 10.1016/j.mad.2022.111726. Epub 2022 Aug 23. PMID: 35998821.
Nash A, Aumann TD, Pigoni M, Lichtenthaler SF, Takeshima H, Munro KM, Gunnersen JM. Lack of Sez6 Family Proteins Impairs Motor Functions, Short-Term Memory, and Cognitive Flexibility and Alters Dendritic Spine Properties. Cereb Cortex. 2020 Apr 14;30(4):2167-2184. doi: 10.1093/cercor/bhz230. PMID: 31711114.

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.