title: Cryopreservation of spleen and lymph nodes as a source of mononuclear cells to be used for the development of monoclonal antibody producing hybridoma cells.
abstract: In the present study, spleen and lymph nodes of mice were cryopreserved as a whole tissue and after thawing, membrane integrity of mononuclear cells was determined by trypan blue exclusion and PI staining. T and B lymphocytes, macrophages and dendritic cells have been isolated from both cryopreserved tissue and analyzed by Flow cytometry. BALB/c mice were immunized with Hepatitis e antigen (HBeAg) and spleen and lymph nodes of mice were cryopreserved for 3 to 10 months. The cells obtained from both tissue were applied to hybridoma technology to understand if the cells keep their viability and functionality. The cells were isolated and fused with F0 mouse myeloma cells and several antibody producing hybrid cells were developed. Results have shown that cryopreserved spleen and lymph nodes of mice can be efficiently used in hybridoma technology for the successful generation of monoclonal antibody producing hybrid cells.
authors.affiliation: TUBITAK Marmara Research Center, Genetic Engineering and Biotechnology Institute. Gebze, Kocaeli, Turkey.
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Sogut
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Hatipoglu
authors.forename: Muge
authors.initials: M
authors.lastname: Serhatli
authors.forename: Safak Isil
authors.initials: SI
authors.lastname: Cevik
authors.forename: Deniz
authors.initials: D
authors.lastname: Durali
authors.forename: Aynur
authors.initials: A
authors.lastname: Basalp
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D015925:Cryopreservation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006825:Hybridomas / Q000166:cytology*; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000502:physiology; D008198:Lymph Nodes / Q000166:cytology*; D051379:Mice; D013154:Spleen / Q000166:cytology*
chemical_list: D000911:Antibodies, Monoclonal