B-LCL-CDG7 Cells
General information
Description | B-LCL-CDG7 is an EBV-transformed B lymphocyte cell line derived from a young boy 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 Beta-mannosidase II or Nacetylglucosaminyltransferase II. |
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Organism | Human |
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. |
Characteristics
Age | Child |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Round cells |
Cell type | B lymphocyte |
Growth properties | Suspension, Cluster |
Identifiers / Biosafety / Citation
Citation | B-LCL-CDG7 (Cytion catalog number 302018) |
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Biosafety level | 2 |
Expression / Mutation
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)+ |
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Antigen expression | CD19+, CD20+, CD37+, CD43+, CD44+, CD45+, CD45R0-MHC Class.I+, MHC Class II (HLA-DR)+ |
Handling
Culture Medium | RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) |
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Medium 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 10^5 cells/ml and keep the cell concentration within the range of 1 x 10^5 to 5 x 10^5 cells/ml for optimal growth. |
Fluid renewal | Once the medium colour turned into yellow |
Freezing recovery | Medium |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
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. |
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STR profile |
Amelogenin: x,y
CSF1PO: 11
D13S317: 12, 14
D16S539: 10, 12
D5S818: 11, 12
D7S820: 8, 10
TH01: 6, 7
TPOX: 8, 11
vWA: 17, 18
D3S1358: 17, 18
D21S11: 30
D18S51: 13, 16
Penta E: 7, 12
Penta D: 9, 14
D8S1179: 11, 13
FGA: 21, 24
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HLA alleles |
A*: 01:01:01, 11:01:01
B*: 35:01:01, 51:01:01
C*: 01:02:01, 04:01:01
DRB1*: 07:01:01, 09:01:02G
DQA1*: 02:01:01, 03:02:01
DQB1*: 02:02:01, 03:03:02
DPB1*: 02:01:02G, 04:02:01G
E: 01:01:01
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