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B-LCL-HROC285
Tissue
Peripheral blood
Disease
Adenocarcinoma
Metastatic site
Not applicable (EBV-transformed B-LCL from Lynch syndrome CRC patient)
Synonyms
B-LCL CO285, Bc HROC285

Documents

B-LCL-HROC285

Cell line · Product number 300869

B-LCL-HROC285 is an Epstein-Barr virus (EBV) transformed B lymphocyte cell line derived from a patient, who had colon adenocarcinoma associated with Lynch...

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01General information

Description

B-LCL-HROC285 is an Epstein-Barr virus (EBV) transformed B lymphocyte cell line derived from a patient, who had colon adenocarcinoma associated with Lynch syndrome. This specific type of colon cancer is linked to hereditary non-polyposis colorectal cancer (HNPCC), commonly caused by mutations in DNA mismatch repair genes. The B-LCL-HROC285 cell line allows for the study of EBV-related transformation processes in B cells, as well as insights into cancer-related immune responses.

The B-LCL-HROC285, provides a valuable tool for understanding the immune system's interactions with cancer cells, particularly how transformed B cells might interact with the immune environment in colorectal cancers arising from Lynch syndrome. This cell line is useful for immunological and oncological studies due to its genetic background and the EBV transformation process, which is known to influence B-cell proliferation and clonal selection.

Tissue
Peripheral blood
Disease
Adenocarcinoma
Metastatic site
Not applicable (EBV-transformed B-LCL from Lynch syndrome CRC patient)
Applications
T cell and NK cell assays; HLA typing; Lynch syndrome immunology; mismatch repair (MMR) deficiency-associated immune response; CTL assay target cells; HROC biobank patient-matched studies
Synonyms
  • B-LCL CO285
  • Bc HROC285

02Classification

Organism
Human
Sex
Female
Growth properties
Suspension
GMO status
GMO-S2
GMO status details
GMO-S2: This B-LCL contains a stably maintained EBV episome (EBNA-1/-2/-3, LMP-1/-2). EBV is classified as risk group 2; BSL-2 containment required. This classification applies within Germany; regulations may differ elsewhere.
Biosafety level
2

03Characteristics

Morphology
Round cells
Cell type
B lymphoblast

04Identifiers

Citation
B-LCL-HROC285 (Cytion catalog number 300869)
NCBI TaxID
9606
Cellosaurus
Not assigned

05Expression and susceptibility

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
Gently homogenize the cell suspension in the flask by pipetting up and down, then take a representative sample to determine the cell density per ml. Dilute the suspension to achieve a cell concentration of 1 x 105 cells/ml with fresh culture medium, and aliquot the adjusted suspension into new flasks for further cultivation.
Thawing and culturing
  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.

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
30 years
Ethnicity
Caucasian