title: Non-mitogen containing conditioned medium for hybridoma production and single cell cloning.
abstract: BACKGROUND
  The hybridoma technique is the standard method for production of monoclonal antibodies of interest. However, the newly formed hybridomas and cells at low density often grow poorly or die. This is the major obstacle to production of monoclonal antibodies.
  
  
  OBJECTIVE
  The aim of this study was to establish a method for preparation of conditioned medium in the absence of mitogen for promoting the growth of hybridomas after cell fusion and during single cell cloning.
  
  
  METHODS
  Culture supernatants were obtained from the cultures of BW5147 mouse thymoma cells without mitogen stimulation. Novel conditioned mediums were investigated for their ability to support hybridoma single-cell cloning and hybridoma production.
  
  
  RESULTS
  We demonstrated that these conditioned mediums could support hybridoma single-cell growth, both for stable and newly generated hybridomas, at a level equal to the commercial conditioned medium BM-Condimed H1. The conditioned medium was most effective at a final concentration of 20% with the basal medium supplemented with 10% fetal calf serum. The novel conditioned medium could also be effectively employed for generation of hybridomas secreting various monoclonal antibodies. Interestingly, fibroblast overgrowth in the post-fusion wells was markedly reduced when using the novel conditioned medium, as compared to the commercial BM-Condimed H1, likely due to the absence of mitogen in the conditioned medium.
  
  
  CONCLUSIONS
  We describe the method for production of a novel conditioned medium for hybridoma technology. This method is simple, needing no special technology or sophisticated equipment. The novel conditioned medium is, therefore, recommended for use in place of expensive commercial conditioned medium for hybridoma technology, especially in resource-limited countries.
authors: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
authors: Napaporn
authors: N
authors: Apiratmateekul
authors: Supansa
authors: S
authors: Pata
authors: Sawitree
authors: S
authors: Chiampanichayakul
authors: Watchara
authors: W
authors: Kasinrerk
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002459:Cell Fusion; D002999:Clone Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006825:Hybridomas / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008934:Mitogens / Q000172:deficiency; D059010:Single-Cell Analysis; D013945:Thymoma / Q000378:metabolism / Q000473:pathology; D013953:Thymus Neoplasms / Q000378:metabolism / Q000473:pathology
chemical_list: D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D008934:Mitogens