KC-5317

CT26-CD13 Cell Line

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Background of CT26-CD13 Cell Line

a receptor for coronaviruses. CD13 functions in cell surface antigen expression by trimming the N-terminal amino acids of MHC class II-bound peptides. CD13 is also a myeloid antigen that can be used for diagnosing acute myeloid leukemia and granulocytic sarcoma. CD13 can also be used to distinguish Down syndrome-related acute myeloid leukemia from transient myeloproliferative disorders.

Specifications

Catalog Number	KC-5317
Cell Line Name	CT26-CD13 Cell Line
Host Cell Line	CT26
Description	Stable CT26 clone expressing exogenous CD13 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 1:4~1:10 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

Cell Line Generation

CT26-CD13 Cell Line was generated using lentivirus expressing cynomolgus CD13 gene sequence.

Characterization

Figure 1: Characterization of CD13 overexpression in the CT26-CD13 stable clone using FACS.

Figure 2: Characterization of CD13 overexpression in the CT26-CD13 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10ug/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 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 9.0mL 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 cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:4 ~ 1:10 every 2~3 days; seed out at about 1-3 x 10 ⁵ 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. Lu C, Amin MA, Fox DA. CD13/Aminopeptidase N Is a Potential Therapeutic Target for Inflammatory Disorders. J Immunol. 2020 Jan 1;204(1):3-11. doi:10.4049/jimmunol.1900868. PMID: 31848300; PMCID: PMC6997018.
2.Guo Q, Li X, Cui MN, Sun JL, Ji HY, Ni BB, Yan MX. CD13: A Key Player in Multidrug Resistance in Cancer Chemotherapy. Oncol Res. 2020 Dec10;28(5):533-540. doi: 10.3727/096504020X15919605976853. Epub 2020 Jun 12. PMID:32532363; PMCID: PMC7751223.
3.Ghosh M, Kelava T, Madunic IV, Kalajzic I, Shapiro LH. CD13 is a criticalregulator of cell-cell fusion in osteoclastogenesis. Sci Rep. 2021 May24;11(1):10736. doi: 10.1038/s41598-021-90271-x. PMID: 34031489; PMCID:PMC8144195.