B-LCL-CDG1

B-LCL-CDG1 is an EBV-transformed B lymphocyte cell line derived from a patient diagnosed with PMM2-CDG, a congenital disorder of glycosylation (CDG). This rare metabolic disorder arises from mutations in the *PMM2* gene, which encodes phosphomannomutase 2 (PMM2), an essential enzyme in the glycosylation pathway. Mutations in *PMM2* disrupt the synthesis of glycosylated oligosaccharide chains, leading to defective glycosylation of various glycoproteins and glycosphingolipids in tissues and blood. The disorder is characterized by multisystemic manifestations, often affecting neurological, hepatic, and endocrine functions.

As an EBV-transformed lymphoblastoid cell line, B-LCL-CDG1 provides a valuable in vitro model for studying the molecular and cellular consequences of *PMM2* deficiency. This cell line can be used to investigate glycosylation defects, PMM2 enzyme activity, and potential therapeutic interventions, including gene correction and substrate supplementation. B-LCL-CDG1, alongside other CDG patient-derived cell lines, serves as a crucial resource for understanding the pathophysiology of CDGs and evaluating novel treatment strategies for these disorders.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

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.

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.

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.