CH12.LX plasmid vector質(zhì)粒載體

Na?ve B-cells differentiate and expand upon antigenic stimulation, undergoing class-switch recombination of immunoglobulin genes and forming antibody-producing plasma cells and memory cells that generate upon recall stimulation (1). B-cell models are useful for elucidating pathways of differentiation and proliferation mechanisms of lymphoma.
The CH12.LX mouse lymphoma cell line is an established and well-characterized B-cell model. CH12.LX cells express Ly-12 and have the characteristic IgM+CD5+CD23-cell surface marker expression of B1a B lymphocytes, as well as expressing CD19, CD11b, MHC II and alpha-4-integrin (3). CH12.LX cells are responsive to a variety of antigens including lipopolysaccharide, expressing the activation protein CD14 (4). CH12.LX cells proliferate rapidly and are ideal for studies of B-cell activation and expansion.

Source:
The CH12.LX cell line was generated from B10.H-2aH4bp/Wts mice immunized with sheep erythrocytes (2).

Research Category:
Cancer
Cell Line Origin
Mouse, Cancer Cells
Packaging
1X106 cells/vial
Storage and Stability
Store in liquid nitrogen. The cells can be cultured for at least 10 passages after initial thawing without significantly affecting the cell marker expression and functionality.

Quality Control Testing
? Each vial contains ≥ 1X106 viable cells.
? Cells are tested negative for infectious diseases by a Mouse
Essential CLEAR panel by Charles River Animal Diagnostic
Services.
? Cells are verified to be of mouse origin and negative for interspecies contamination from rat, human, chinese hamster, Golden
Syrian hamster, and non-human primate (NHP) as assessed by a
Contamination CLEAR panel by Charles River Animal Diagnostic
Services.
? Cells are negative for mycoplasma contamination

Image:

Figure 1. Brightfield image of CH12.LX mouse B-cell lymphoma
cells.


RPMI-1640 +10%FBS+ 100 U/ml penicillin G sodium (option); 100 μg/ml streptomycin sulfate (option);

Complete growth medium, 95%; DMSO, 5%


Protocols
CH12.LX Mouse Lymphoma cell line grow as suspension cells and thus do not require enzymatic detachment or dissociation.
Passage when the cell density reaches 1–1.5 million cells/mL. Optimal plating density should be ~200,000 - 250,000 cells/mL. The cells should not be grown at excessively high densities.
Important Note: CH12.LX cells grow relatively slowly and may take several days to reach optimal density for passaging.
1. Do not thaw the cells until the recommended medium is on hand. Cells are thawed in RPMI-1640 (Sigma Cat. No. R0883)
supplemented with 2 mM Glutamine and 20% FBS. Once established, cells are expanded in RPMI-1640 with 2 mM glutamine and 10% FBS.
2. Remove the vial of frozen CH12.LX cells from liquid nitrogen and incubate in a 37°C water bath. Closely monitor until the cells are completely thawed. Maximum cell viability is dependent on the rapid and complete thawing of frozen cells.
IMPORTANT: Do not vortex the cells.
3. As soon as the cells are completely thawed, disinfect the outside of the vial with 70% ethanol. Proceed immediately to the next step.
4. In a laminar flow hood, use a 1 or 2 mL pipette to transfer the cells to a sterile 15 mL conical tube. Be careful not to introduce any bubbles during the transfer process.
5. Using a 10 mL pipette, slowly add dropwise 9 mL of CH12.LX Thaw Medium (Step 1 above) to the 15 mL conical tube.
IMPORTANT: Do not add the entire volume of media all at once to the cells. This may result in decreased cell viability due
to osmotic shock.
6. Gently mix the cell suspension by slowly pipetting up and down twice. Be careful not to introduce any bubbles.
IMPORTANT: Do not vortex the cells.
7. Centrifuge the tube at 300 x g for 2-3 minutes to pellet the cells.
8. Decant as much of the supernatant as possible. Steps 5-8 are necessary to remove residual cryopreservative (DMSO).
9. Resuspend the cells in 15–20 mL of CH12.LX Thaw Medium.
10. Transfer the cell suspension to a T75 flask.
11. Incubate the cells at 37°C in a humidified incubator with 5% CO2. CH12.LX suspension cells require media replenishment every 2-3 days. Once established, CH12.LX cells are expanded in RPMI-1640 with 2 mM glutamine and 10% FBS. Passage cells when the cell density is at 1 -1.5 million cells/mL.
12. Cells are typically plated at a density of 200,000 - 250,000 cells/mL

Cryopreservation of Cells
CH12.LX Mouse B-Cell Lymphoma Cell Line may be frozen in the expansion medium plus 10% DMSO using a Nalgene slow freeze Mr. Frosty container.

Supplier來(lái)源:BioVector NTCC Inc.
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