KC-3984

Jurkat-NFAT-Luc2-CD7-KO-3A4 Cell Line

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Background of Jurkat-NFAT-Luc2-CD7-KO-3A4 Cell Line

CD7 is located on chromosome 17 and encodes a cell surface glycoprotein member of the immunoglobulin superfamily. The protein is found on thymocytes and mature NK and T-cells. It plays an essential role in T-cell interactions and also in T-cell/B-cell interactions during early lymphopoeisis. CD7 is highly expressed on the surface of T-ALL/LBL T cells and is considered a viable CAR-T therapeutic target. Although CD7 is expressed on natural killer (NK) cells and normal T cells at various stages of maturation, CD7 knockout mice develop normal lymphoid organs and immune responses, suggesting that CD7 is dispensable for T/NK cell development and functions.

Specifications

Catalog Number	KC-3984
Cell Line Name	Jurkat-NFAT-Luc2-CD7-KO-3A4 Cell Line
Host Cell Line	Jurkat-NFAT-Luc2
Description	Stable Jurkat-NFAT-Luc2-reporter cell line with CD7 gene knockout
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+300μg/ml Hygromycin B
Selection Marker	NA
Morphology	Lymphoblast
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

Jurkat-NFAT-Luc2-CD7-KO-3A4 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Jurkat-NFAT-Luc2-CD7-KO Cell Line stable clone using FACS.

Figure 2: Characterization of Jurkat-NFAT-Luc2-CD7-KO-3A4 cell line stable clone using PCR sequencing.

Figure 3: Characterization of Jurkat-NFAT-Luc2-CD7-KO-3A4 cell line stable clone using RT-PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS+300μg/ml Hygromycin B)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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 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

Rogers SL, Zhao Y, Jiang X, Eaves CJ, Mager DL, Rouhi A. Expression of the leukemic prognostic marker CD7 is linked to epigenetic modifications in chronic myeloid leukemia. Mol Cancer. 2010 Feb 22;9:41. doi: 10.1186/1476-4598-9-41. PMID: 20175919; PMCID: PMC2843654.
Lu P, Liu Y, Yang J, Zhang X, Yang X, Wang H, Wang L, Wang Q, Jin D, Li J, Huang X. Naturally selected CD7 CAR-T therapy without genetic manipulations for T-ALL/LBL: first-in-human phase 1 clinical trial. Blood. 2022 Jul 28;140(4):321-334. doi: 10.1182/blood.2021014498. PMID: 35500125.