KC-4498

A549-ROR1-High Cell Line

54923

Home » A549-ROR1-High Cell Line

Background of A549-ROR1-High Cell Line

ROR1, also named as NTRKR1, is a transmembrane protein belong to the receptor tyrosine kinase-like orphan receptor (ROR) family. ROR1 modulates neurite growth in the central nervous system. ROR1 has recently found to be overexpressed on cancer stem cell, which play a function role in promoting cancer cell migration, invasion or spheroid formation.

Specifications

Catalog Number	KC-4498
Cell Line Name	A549-ROR1-High Cell Line
Clone Number	2-1#
Host Cell Line	A549
Description	Stable A549 clone expressing exogenous ROR1 gene in high level
Quantity	One vial 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	Epithelial
Subculture	Split saturated culture 1:6-1:8 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 18 hours
Mycoplasma Status	Negative

Cell Line Generation

A549 ROR1 high cell line was generated using a lentiviral vector expressing the ROR1 sequence.

Characterization

Figure 1: Characterization of ROR1 overexpression in the A459 ROR1 stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 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 1:6-1:8 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

Masiakowski P, Carroll RD (Dec 1992). A novel family of cell surface receptors with tyrosine kinase-like domain. The Journal of Biological Chemistry. 267 (36): 26181–90.2. Reddy UR, Phatak S, Allen C, Nycum LM, Sulman EP, White PS, Biegel JA (Apr 1997). Localization of the humanRor1 gene (NTRKR1) to chromosome 1p31-p32 by fluorescence in situ hybridization and somatic cell hybridanalysis. Genomics. 41 (2): 283–5.3.
Baskar S, Kwong KY, Hofer T, Levy JM, Kennedy MG, Lee E, Staudt LM, Wilson WH, Wiestner A, Rader C (Jan2008). Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocyticleukemia. Clinical Cancer Research. 14(2): 396–404.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.