KC-1367

RS4;11-BCL2-F104L-Cell-Line

22003

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Background of RS4;11-BCL2-F104L-Cell-Line

Bcl-2 (B-cell lymphoma 2), as it is the second member of a range of proteins initially described in chromosomal translocations involving chromosomes 14 and 18 in follicular lymphomas. Bcl-2-family proteins regulate all major types of cell death, including apoptosis, necrosis and autophagy, thus operating as nodal points at the convergence of multiple pathways with broad relevance to oncology. Venetoclax is the first bcl-2 protein inhibitor approved by FDA. In recent years, bcl-2 small molecule inhibitors (such as ABT-737, ABT -263, ABT - 199 and GX-15-070) have entered clinical trials. RS4;11. a lymphoblast cell line of human. This line was established from the bone marrow of a patient with acute lymphoblastic leukemia. The cells lack surface and cytoplasmic immunoglobulin, and are negative for CALLA (CD10).

Specifications

Catalog Number	KC-1367
Cell Line Name	RS4;11-BCL2-F104L-Cell-Line
Description	Stable RS4;11 expressing exogenous BCL2 gene bearing F104L amino acid mutation.
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	RPMI1640+10%FBS+1µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture 1:2-1:4 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 50 hours
Mycoplasma Status	Negative
In Vivo Validation	Yes

Cell Line Generation

RS4;11 BCL2-F104L cell line was generated using retrovirus vector expressing human BCL2 sequence with F104L mutation and N-terminal Flag tag.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 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 1:2-1:4 every 2-3 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

K W Yip & J C Reed, Bcl-2 family proteins and cancer. Oncogene volume 27, pages 6398ÿ6406 (27 October 2008)