KC-1336

CHOK1-mouse-CD47-Cell-Line

21947

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Background of CHOK1-mouse-CD47-Cell-Line

CD47 (originally named integrin-associated protein (IAP)) is a cell surface protein of the immunoglobulin (Ig) superfamily, which is heavily glycosylated and expressed by virtually all cells in the body. plays a pivotal role in this balance by delivering a "don't eat me signal" upon binding to the Signal-regulatory protein alpha (SIRPα) receptor on myeloid cells. and is potential therapeutic target in some cancer and treatment of pulmonary fibrosis. Diseases associated with CD47 include Hereditary Spherocytosis and Hemolytic Anemia.

Specifications

Catalog Number	KC-1336
Cell Line Name	CHOK1-mouse-CD47-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse CD47 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 CD47 Cell Line was generated using a lentiviral vector expressing the mouse CD47 sequence.

Characterization

Figure 1: Characterization of mouse CD47 overexpression in the CHO-K1 mouse CD47 stable clone 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

Oldenborg PA. CD47: A Cell Surface Glycoprotein Which Regulates Multiple Functions of Hematopoietic Cells in Health and Disease. ISRN Hematol. 2013;2013:614619. doi: 10.1155/2013/614619. Epub 2013 Jan 21. PMID: 23401787; PMCID: PMC3564380.
Jie XL, Kong YY, Zhou GB. [Latest Findings on the Role of CD47 in Tumor Immune Evasion and Related Targeted Therapies]. Sichuan Da Xue Xue Bao Yi Xue Ban. 2023 May;54(3):455-461. Chinese. doi: 10.12182/20230560101. PMID: 37248568; PMCID: PMC10475431.
Fenalti, G., Villanueva, N., Griffith, M. et al. Structure of the human marker of self 5-transmembrane receptor CD47. Nat Commun 12, 5218 (2021).