The aim of this work was to investigate the minimum technical requirements for production of
live offspring with somatic cell nuclear transfer. The experiment was performed in a field type
laboratory without micromanipulators and carbon dioxide incubators. All long-term incubations
were performed in the Submarine Incubation System (SIS) using various gasmixtures. The somatic
cell culture was established from ear biopsy of a 9-year-old Holstein cow. Nuclear transfer was
performed using the Handmade Cloning (HMC) technique. Zona-free oocytes were randomly
bisected by hand with a disposable blade and a stereomicroscope. Cytoplast were selected using
Hoechst staining and a fluorescent microscope. After a two-step fusion embryos were activated
with calcium ionophore and dimethylaminopurine. Embryos were cultured in microwells (WOWs)
in SOFaaci medium supplemented with 5% cattle serum. In two consecutive experiments, six
blastocysts were produced from 52 reconstructed embryos. On Day 7, five blastocysts were
transferred into synchronized recipients. All three recipients became pregnant but two pregnancies
aborted at 6 and 7 months, respectively. A heifer calf weighing 27 kg was delivered at term by
Caesarean section from the third pregnancy. The healthy 6-month-old heifer, the first cloned animalof Africa, is living evidence that nuclear transfer technology may be successfully used under basic
laboratory conditions.