KC-6385

293T-cyno-CD98HC Cell Line

62097

Home » 293T-cyno-CD98HC Cell Line

Background of 293T-cyno-CD98HC Cell Line

CD98 heavy chain (CD98hc) forms heteromeric amino acid (AA) transporters by interacting with different light chains. Cancer cells overexpress CD98hc-transporters in order to meet their increased nutritional and antioxidant demands, since they provide branched-chain AA (BCAA) and aromatic AA (AAA) availability while protecting cells from oxidative stress. Involved in L-leucine import across plasma membrane. Located in cell cortex. Is expressed in several structures, including adult head; adult heart; embryonic/larval midgut primordium; ganglia; and gut section. Human ortholog(s) of this gene implicated in lung non-small cell carcinoma. Orthologous to human SLC3A2 (solute carrier family 3 member 2).

Specifications

Catalog Number	KC-6385
Cell Line Name	293T-cyno-CD98HC Cell Line
NCBI/UniProt Accession Number	XM_045372009.2
Clone Number	5#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous cyno CD98HC 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:4-1:5 every 2-3 days; seed out at about 1-3 x 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 28 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-cyno-CD98HC cell line was generated using a lentiviral vector expressing the cyno CD98HC sequence.

Characterization

Figure 1: Characterization of cyno CD98HC overexpression in the 293T-cyno-CD98HC stable clone using PCR sequencing.

Figure 2: Characterization of cyno CD98HC overexpression in the 293T-cyno-CD98HC stable clone using FACS.

Cell Resuscitation

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

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Vection S, O\'Callaghan D, Keriel A. CD98hc in host-pathogen interactions: roles of the multifunctional host protein during infections. FEMS Microbiol Rev. 2022 Sep 2;46(5):fuac023.
Cano-Crespo S, Chillarón J, Junza A, Fernández-Miranda G, García J, Polte C, R de la Ballina L, Ignatova Z, Yanes Ó, Zorzano A, Stephan-Otto Attolini C, Palacín M. CD98hc (SLC3A2) sustains amino acid and nucleotide availability for cell cycle progression. Sci Rep. 2019 Oct 1;9(1):14065.