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The Use of Bull Round Spermatids for Producing Reconstructed Embryos
Recently, sperm has been used as a vector to carry exogenous genes for the production of transgenic animals. However, the success in cattle is low, due to deficiencies in oocyte activation and sperm decondensation caused by high disulphide bond (S=S) content in mature sperm. This study was carried out to develop an effective method for producing transgenic animals with round spermatids (RS). Two methods of embryo production - electric fusion (EC) or intracyto-plasmic injection (IC) and three activation treatments were compared. RS were isolated from bull testes by Percoll density gradients (20, 35, 40, 45 and 90%). Fusion between ooplast and RS was performed with a single DC electric pulse (1.0 KV/cm, 45 sec) in 0.28 M mannitol solution supplemented with 100 M CaCl2 and 100 M MgCl. (중략)
