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B-LCL-CDG4
Cell line · Product number 302015
B-LCL-CDG4 is an EBV-transformed B lymphocyte cell line derived from a young girl with CDAII. CDAII is a rare genetic anaemia, affiliated to the class of CDG...
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01General information
- Description
- B-LCL-CDG4 is an EBV-transformed B lymphocyte cell line derived from a young girl with CDAII. CDAII is a rare genetic anaemia, affiliated to the class of CDG glycosylation disorders. CDAII patients have a defect in the COPII component SEC23B gene which is involved in the intracellular protein transport system (in particular vesicular budding from ER). The respective patient is homozygous for the mutation in this gene. Band 3 glycoprotein of erythrocyte membranes is under glycosylated by aberrant glycosylation of polylactosamine motifs of glycoproteins but not of glycosphingolipids, thus band 3 of CDA II erythrocytes have truncated hybrid-type oligosaccharides. This points to an additional defect in the Golgi glycosylation enzymes mannosidase II or Nacetylglucosaminyltransferase II.
- Tissue
- Peripheral blood
- Disease
- Congenital Disorders of Glycosylation
- Applications
- Genotyping of CDG effects in immune cells, functional testing (e.g. B cell surface antigens), testing of cytotoxic drugs, mutational analysis, analysis of apoptotic mechanisms, HLA-typing, impact of defective glycosylation of distinct cellular glycoproteins on diverse functions.
02Classification
- Organism
- Human
- Sex
- Female
- Growth properties
- Suspension
- Biosafety level
- 2
03Characteristics
- Morphology
- Round cells
- Cell type
- B lymphocyte
04Identifiers
- Citation
- B-LCL-CDG4 (Cytion catalog number 302015)
- NCBI TaxID
- 9606
- Cellosaurus
- CVCL_A9Y2
05Expression and susceptibility
- Surface antigens
- CD15 (Lewis x)+, CD15s (sialylated Lewis x)-, CD75s (sialylated lactosaminyl Noligosccharides)+, CD173 (blood group H)-, CD174 (blood group Lewis y)-, CD175 (Tn)-, CD175s (sialylated Tn)-, CD176 (TF)+
- Antigen expression
- CD19+, CD20+, CD37+, CD43+, CD44+, CD45+, CD45R0-, MHC Cl.I+, MHC Class II (HLA-DR)+
- Viruses
- Transformant: EBV
06Handling and culture
- Culture medium
- RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- Incubation atmosphere
37°C, 5% CO2, humidified atmosphere.
- Supplements
- Supplement the medium with 10% heat-inactivated FBS
- Subculturing
- Maintain cultures by periodically adding or replacing the medium. Initiate cultures with a density of 2 x 105 cells/ml and keep the cell concentration within the range of 3 x 105 to 5 x 105 cells/ml for optimal growth.
- Fluid renewal
- Once the medium colour turned into yellow
- Post-thaw recovery
- Medium
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.
For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.
Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.
Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
07Quality control and cryopreservation
- Sterility
Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.
To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.
- Freeze medium
- As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
- Storage conditions
For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.
- Shipping conditions
Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.
08Donor
- Donor age
- Child
- Ethnicity
- Caucasian
09Legal and safety
- Safety precautions
- When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.
- Warranty
At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.
For full details, please refer to our Warranty Statement.
- Material Transfer Agreement required
- Yes