KC-4936

PA-1-ROR1-KO Cell Line

53099

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Background of PA-1-ROR1-KO Cell Line

The ROR1 gene, encoding the Receptor Tyrosine Kinase Like Orphan Receptor 1, plays a critical role in embryonic development and has been implicated in various cancers. ROR1 is a transmembrane protein that belongs to the receptor tyrosine kinase family, which is essential for cell proliferation, differentiation, and survival. During embryogenesis, ROR1 is highly expressed and involved in the regulation of skeletal and neuronal development. However, its expression is significantly downregulated in most adult tissues. Aberrant expression of ROR1 has been observed in several malignancies, including chronic lymphocytic leukemia (CLL), breast cancer, and lung cancer, where it promotes tumor growth, survival, and metastasis.

Specifications

Catalog Number	KC-4936
Cell Line Name	PA-1-ROR1-KO Cell Line
Host Cell Line	PA-1
Description	Stable PA-1 cell line with ROR1 gene knockout, No.5C4
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
Selection Marker	NA
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 36 hours
Mycoplasma Status	Negative

Cell Line Generation

PA-1-ROR1-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of PA-1-ROR1-KO Cell Line stable clone using PCR sequencing.

Figure 2: Characterization of PA-1-ROR1-KO Cell Line stable clone using RT-PCR sequencing.

Figure 3: Characterization of PA-1-ROR1-KO Cell Line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM + 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 1:4-1:10 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

Zhang S, Chen L, Wang-Rodriguez J, Zhang L, Cui B, Frankel W, Wu R, Kipps TJ. The onco-embryonic antigen ROR1 is expressed by a variety of human cancers. Am J Pathol. 2012 Dec;181(6):1903-10. doi: 10.1016/j.ajpath.2012.08.024. Epub 2012 Oct 4. PMID: 23041612; PMCID: PMC3509760.
Fukuda T, Chen L, Endo T, Tang L, Lu D, Castro JE, Widhopf GF 2nd, Rassenti LZ, Cantwell MJ, Prussak CE, Carson DA, Kipps TJ. Antisera induced by infusions of autologous Ad-CD154-leukemia B cells identify ROR1 as an oncofetal antigen and receptor for Wnt5a. Proc Natl Acad Sci U S A. 2008 Feb 26;105(8):3047-52. doi: 10.1073/pnas.0712148105. Epub 2008 Feb 19. PMID: 18287027; PMCID: PMC2268582.