KC-2404

B16/F10-ROR1 Cell Line

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Background

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-2404
Cell Line Name:
B16/F10 ROR1 Cell Line
Host Cell Line:
Mouse B16F10 cell line
Description:
Stable B16/F10 clone expressing exogenous mouse ROR1
Quantity:
One vial of frozen cells (5X106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% DMEM+20% FBS+10% DMSO
Propagation Medium:
DMEM+10%FBS+1μg/mL Puromycin
Selection Marker:
Puromycin
Morphology:
Epithelial
Subculture:
Split saturated culture 1:4-1:10 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 17 hours
Mycoplasma Status:
Negative

Cell Line Generation

B16F10 human ROR1 Cell Line was generated using a lentiviral vector expressing the human ROR1 sequence.

Characterization

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Application

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Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS + 1μg/mL Puromycin) in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO2 incubator. 8. Split saturated culture 1:4-1:10 every 2-3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage

References

1. 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 human Ror1 gene (NTRKR1) to chromosome 1p31-p32 by fluorescence in situ hybridization and somatic cell hybrid analysis". 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 (Jan 2008). "Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocytic leukemia". Clinical Cancer Research. 14(2): 396–404.
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