CL-01 Cell Line

Product Discontinued

Applications/Dilutions

• Application Notes: NBP1-49595 is useful to study the mechanism of B cell differentiation and survival in vitro during antibody responses. It is also useful to study gene regulation, function, and cellular and molecular events in B cells under normal or pathological conditions. THIS CELL LINE IS ONLY AVAILABLE TO NOT-FOR-PROFIT ACADEMIC INSTITUTIONS OR COMMERCIAL ENTITIES THAT HAVE SECURED A SEPERATE COMMERCIAL USE LICENSE. Please email monica@novusbio.com for more information.

Packaging, Storage & Formulations

• Storage: Store in gas phase of liquid nitrogen.
• Buffer: Approximately 3x10 ^ 6 cells in 1.5ml of freezing media and 10% DMSO.

Notes

This cell line is a human B cell line with IgM+ IgD+ on the surface. However, this is also EBV transformed, so the cells can activate each other easily by cell-cell contact. The cells will not be IgM+IgD+ once they get activated and class switched. It is recommended to keep the cells in low density to prevent the activation, but be sure to avoid too low of a density because the cells will die. It is recommended to culture 5-10 mil cells in 50ml medium (T75 flask). However, if you don't care about the activation, for example, you will want to get mRNA or gDNA or proteins for molecular or biochemistry work, it is fine to increase the density to 0.5-1Mil/ml.

CULTURE CONDITIONS:
The first time you thaw the tube, after removing DMSO (spin with low speed, 700 rpm), resuspend the cells in a T75 flask with 30ml complete medium (RPMI 1640, supplemented with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine). Keep the flask horizontal. After two days, generally spin down the cells and remove debris (700rpm, 3 or 5 mins), resuspend the cells in 50ml medium until you have enough cells to freeze back. Change the medium 2-3 days by simply removing some supernatant and adding some fresh medium.

The cell density should be controlled within the range between 0.1-0.2 mil/ml. They will be hyper-activated if too crowded, and die if too diluted.

Freeze as many vials as you can from the beginning (around 5-8 vials). Each time you thaw one vial, freeze back 2-3 vials, culture the rest of the cells for your experiments, and discard all the cells after 4-5 generations. For new experiments, thaw a new vial.

Alternate Names for CL-01 Cell Line

• CL01 cell line
• CL01 cells
• CL-01 cells
• Human Monoclonal IgM+IgD+ B Cell Line

Background

CL-01 cell line is the only non-antigen specific human monoclonal B cell line that expresses surface IgM and IgD, therefore ready to differentiate upon appropriate stimuli. CL-01 cells are physiological stimuli inducible and can display the phenotypic changes putatively attributed to B cells during maturation. CL-01 cells can undergo somatic hypermutation, Ig class switching and complete differentiation in vitro. Compared to previously used polyclonal cells, CL-01 cells are available in large amounts; they are homogenous in genetic makeup and achieve higher switching efficiency. These features make the CL-01 cell line an ideal model system for in vitro studies of human B cell differentiation and for cancer, autoimmune, and infectious diseases drug development.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are guaranteed for 6 months from date of receipt.

Publications for CL-01 Cell Line (NBP1-49595)(3)

Publications using NBP1-49595

• Schmidt C, Christian L, Smith T et al. Lipid Rafts Interaction of the ARID3A Transcription Factor with EZRIN and G-Actin Regulates B-Cell Receptor Signaling Diseases 2021-03-20 [PMID: 33804610]
• Bernstein RM, Mills FC, Mitchell M, Max EE. Complex mechanisms for inhibition of immunoglobulin gene expression in a germinal center B cell line. Mol Immunol;41(1):63-72. 2004-05-01 [PMID: 15140576]
• Cerutti A, Zan H, Schaffer A, Bergsagel L, Harindranath N, Max EE, Casali P. CD40 ligand and appropriate cytokines induce switching to IgG, IgA, and IgE and coordinated germinal center and plasmacytoid phenotypic differentiation in a human monoclonal IgM+IgD+ B cell line. J Immunol;160(5):2145-57. 1998-03-01 [PMID: 9498752] (ICC/IF, WB, FLOW, Human)

FAQs for CL-01 Cell Line (NBP1-49595). (Showing 1 - 3 of 3 FAQs).

1. Is the cell line CL-01 EBV-transformed and has the status of EBV been analyzed, i.e. can it be lytically induced? Is this cell line an S2 line, i.e. requires biological safety level S2?
   • This cell line is a human B cell line with IgM+ IgD+ on the surface. However, this is also EBV transformed, so the cells can activate each other easily by cell-cell contact. The cells will not be IgM+IgD+ once they get activated and class switched. It should be treated as Biosafety level 1, just as many other human cell lines.
2. I have a question regarding this cell line. Your data sheet states it is transformed with EBV, so does that mean the culture conditions are Cat 2, or has this cell line been shown not to shed virus?
   • Our recommended Biosafety level for our CL-01 cells is Biosafety level 1. The Epstein Barr virus that is used to transform this cell line should not be shed in culture and has not been found to do so in our testing of this cell line.
3. Can you tell me how to maintain the cells ?
   • The first time you thaw the tube, after removing DMSO (spin with low speed, 700 rpm), resuspend the cells in a T75 flask with 30ml complete medium (RPMI 1640, supplemented with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine). Keep the flask horizontal. After two days, generally spin down the cells and remove debris (700rpm, 3 or 5 mins), resuspend the cells in 50ml medium until you have enough cells to freeze back. Change the medium 2-3 days by simply removing some supernatant and adding some fresh medium. The cell density should be controlled within the range between 0.1-0.2 mil/ml. They will be hyper-activated if too crowded, and die if too diluted. Freeze as many vials as you can from the beginning (around 5-8 vials). Each time you thaw one vial, freeze back 2-3 vials, culture the rest of the cells for your experiments, and discard all the cells after 4-5 generations. For new experiments, thaw a new vial.