KC-3953

Ba/F3-human-ROR1-High-Cell-Line

40364

Home » Ba/F3-human-ROR1-High-Cell-Line

Background of Ba/F3-human-ROR1-High-Cell-Line

ROR1 is a membrane receptor that plays a key role in development. It plays an important role in embryonic development by regulating cell migration, cell polarity, neural patterning, and organogenesis. It is highly expressed in the embryonic stage and relatively low in some normal adult tissues. ROR1 is highly expressed in malignant tumors such as leukemia, lymphoma and some solid tumors, which is expected to become a new target for tumor treatment.

Specifications

Catalog Number	KC-3953
Cell Line Name	Ba/F3-human-ROR1-High-Cell-Line
Host Cell Line	Ba/F3
Description	Stable Ba/F3 clone expressing exogenous human ROR1 gene in high level.
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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	1640+10%FBS+1μg/mL puromycin+8ng/mL IL-3
Selection Marker	Puromycin
Morphology	Mostly single, round (some polymorph) cells in suspension
Subculture	Split saturated culture 1:10 every 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 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

BA/F3 human ROR1 high cell line was generated using a lentiviral vector expressing the human ROR1 sequence.

Characterization

Figure 1: Characterization of human-ROR1 overexpression in the Ba/F3-human-ROR1 stable clone using FACS.

Figure 2: Characterization of human-ROR1 in the Ba/F3-human-ROR1 stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS+1μg/mL puromycin+8ng/mL IL-3)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 1:10 every 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

Balakrishnan A, Goodpaster T, Randolph-Habecker J, Hoffstrom BG, Jalikis FG, Koch LK, Berger C, Kosasih PL, Rajan A, Sommermeyer D, Porter PL, Riddell SR. Analysis of ROR1 Protein Expression in Human Cancer and Normal Tissues. Clin Cancer Res. 2017 Jun 15;23(12):3061-3071. doi: 10.1158/1078-0432.CCR-16-2083. Epub 2016 Nov 16. PMID: 27852699; PMCID: PMC5440207.
Kipps TJ. ROR1: an orphan becomes apparent. Blood. 2022 Oct 6;140(14):1583-1591. doi: 10.1182/blood.2021014760. PMID: 35580162; PMCID: PMC10653015.