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Colcemid treatment of myeloma prior to cell fusion increases the yield of hybridomas between myeloma and splenocyte
Authors:M Miyahara  H Nakamura  Y Hamaguchi
Institution:Meiji Institute of Health Science, Naruda, Odawara, Kanagawa 250, Japan
Abstract:Effect of Colcemid treatment of myeloma (X63-Ag8-6.5.3.) prior to fusion with mouse spleen cell was studied in terms of hybridoma formation. Spleen cells from BALB/c mice immunized with various soluble antigens were fused with the myeloma cells by using polyethylene glycol solution. Colcemid treatment of myeloma cells prior to fusion increased the average number of hybridoma colonies per well by 26-570%. The yield of hybridomas producing antigen-specific antibodies was also higher with the Colcemid treatment. The results suggest that most of the proliferative hybridomas are formed by fusion of cells in the M-phase of the cell cycle.
Keywords:ABTS  2  2-azino-di[3-ethylbenzthiazoline sulfonic acid]  ELISA  enzyme-linked immunosorbent assay  HAT  hypoxanthine-aminopterine-thymidine  HCG  human chorionic gonadotropin  i  p    intraperitoneally  IU  international unit  MEM  (Eagle's) minimum essential medium  OPD
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