KC-5479

HPB-ALL-IL-7Ra-KO Cell Line

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Background of HPB-ALL-IL-7Ra-KO Cell Line

The interleukin-7 receptor alpha (IL-7Ra) gene, located on chromosome 5p13 in humans, encodes a critical subunit of the IL-7 receptor (IL-7R), which is essential for lymphocyte development, homeostasis, and immune responses. IL-7Ra pairs with the common gamma chain (γc) to form the functional IL-7R, mediating signals that promote survival, proliferation, and differentiation of T and B cells. Dysregulation of IL-7Ra signaling is associated with severe immunodeficiency, autoimmune diseases, and hematologic malignancies. Single nucleotide polymorphisms (SNPs) in IL-7Ra have been linked to multiple sclerosis (MS) and other autoimmune disorders, highlighting its immunomodulatory significance. Studies using murine models have further demonstrated its non-redundant role in lymphoid development.

Specifications

Catalog Number	KC-5479
Cell Line Name	HPB-ALL-IL-7Ra-KO Cell Line
Clone Number	2C1
Host Cell Line	HPB-ALL
Description	Stable HPB-ALL clone with IL-7Ra gene knockout, No.2C1
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
Selection Marker	NA
Morphology	Lymphoblast
Subculture	Split saturated culture 1:3-1:6 every 1-3 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 50 hours
Mycoplasma Status	Negative

Cell Line Generation

HPB-ALL-IL-7Ra-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of HPB-ALL-IL-7Ra-KO Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of HPB-ALL-IL-7Ra-KO Cell Line stable clone using RT-PCR sequencing.

Figure 3: Characterization of HPB-ALL(Left) and HPB-ALL-IL-7Ra-KO(Right) Cell Lines stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS) 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 1:3-1:6 every 1-3 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

Mazzucchelli R, Durum SK. Interleukin-7 receptor expression: intelligent design. Nat Rev Immunol. 2007 Feb;7(2):144-54. doi: 10.1038/nri2023. PMID: 17259970.
Gregory SG, Schmidt S, Seth P, Oksenberg JR, Hart J, Prokop A, Caillier SJ, Ban M, Goris A, Barcellos LF, Lincoln R, McCauley JL, Sawcer SJ, Compston DA, Dubois B, Hauser SL, Garcia-Blanco MA, Pericak-Vance MA, Haines JL; Multiple Sclerosis Genetics Group. Interleukin 7 receptor alpha chain (IL7R) shows allelic and functional association with multiple sclerosis. Nat Genet. 2007 Sep;39(9):1083-91. doi: 10.1038/ng2103. Epub 2007 Jul 29. PMID: 17660817.