PLH Cells

Categories: B lymphoblast cell lines Description: The PLH cell line is an Epstein-Barr virus (EBV)-transformed human lymphoblastoid cell line derived from a patient with congenital adrenal hyperplasia (CAH) due to steroid 21-hydroxylase (21-OHase) deficiency. This autosomal recessive disorder, which impairs cortisol biosynthesis, is strongly linked to specific HLA haplotypes, particularly HLA-Bw47,DR7. The PLH line is homozygous for this haplotype and has been used as a genetic model to investigate the molecular basis of 21-OHase deficiency. It is especially valuable in studying gene deletions affecting the cytochrome P-450C21 gene, which is responsible for 21-hydroxylation, a crucial step in cortisol production. Molecular analyses using DNA probes confirmed that PLH cells exhibit a homozygous deletion of one of the two P-450C21 genes, consistent with the loss of 21-hydroxylase activity observed in affected individuals. The PLH cell line was part of the Fourth Asia-Oceania Histocompatibility Workshop (4AOHW) panel, which aimed to provide a well-characterized set of EBV-transformed lymphoblastoid cell lines representing diverse MHC alleles and haplotypes. These panels serve as essential resources for histocompatibility studies, HLA typing development, and immunogenetics research. The selection of PLH for inclusion in the 4AOHW reflected its unique MHC genotype and disease relevance, contributing to both the standardization of HLA allele assignments and studies exploring the genetic architecture of immune-related disorders. Organism: Human Tissue: Adrenal gland Disease: Classic congenital adrenal hyperplasia due to 21-hydroxylase deficiency Metastatic Site: Peripheral blood Age: Unspecified Gender: Female Ethnicity: Scandinavian, Caucasian Morphology: Lymphoblast Cell Type: B Cell Growth Properties: Suspension Citation: PLH (Cytion catalog number 302137) Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_E810 Viruses: Epstein-Barr virus (EBV) Culture Medium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) Supplements: Supplement the medium with 10% 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. Freeze Medium: As a cryopreservation medium, 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. Thawing And Culturing Cells: 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. 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. 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: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 1: 820700a Required Product 3: 860015.0 Required Product 4: 830100.0