Production of a cloned calf from a fetal fibroblast cell line

Brazilian Journal of Medical and Biological Research

M.r.b. Mello²  H.v.a. Caetano²  M.g. Marques²  M.s. Padilha²  J.f. Garcia¹  M.p. Milazzotto²  M.e.o.a. Assumpção²  A.s. Lima²  A.c. Nicácio²  C.m. Mendes²  V.p. Oliveira²  J.a. Visintin² 

[1] ,Universidade de São Paulo Faculdade de Medicina Veterinária Departamento de Reprodução AnimalSão Paulo SP ,Brasil

关键词: Cloning;    Nuclear transfer;    Bovine;    Fetal fibroblasts;    Embryo reconstruction;   

DOI  :  10.1590/S0100-879X2003001100006

来源: SciELO

【 摘 要 】

The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5%) were successfully fused and 24 (28.9%) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8%) of which were pregnant on day 35 and 3 (16.7%) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).

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