Conditioning of karyoplasts for producing somatic nuclear transferred gonadal germ cells in domestic chickens.
The objective of this study was to establish a protocol for generating karyoplasts that can be used to produce somatic nuclear transferred gonadal germ cells (snt-GGCs) in domestic chickens. Karyoplasts were produced by centrifuging cultured fibroblasts from 10-day-old chick embryos at 10,000 x g in the presence of 1.0 microg/ml cytochalasin B. The number of karyoplasts was significantly (P<0.05) higher and the diameters of the karyoplasts were significantly (P<0.05) smaller when fibroblasts were centrifuged for 60 min than for 10 or 30 min. It was possible to generate snt-GGCs by electrofusion of GGCs with karyoplasts produced from cryopreserved or serum-starved fibroblasts. These results indicate that karyoplasts generated from 10-day-old chick embryos can be used to produce snt-GGCs even after cryopreservation and serum starvation of the fibroblasts.
Kohara
Yusaku
Y
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan.
Minematsu
Takeo
T
Aikawa
Takeshi
T
Kanai
Yukio
Y
Tajima
Atsushi
A
D000818:Animals; D002467:Cell Nucleus; D002498:Centrifugation; D002642:Chick Embryo; D002645:Chickens; D015925:Cryopreservation; D016895:Culture Media, Serum-Free; D003593:Cytoplasm; D005260:Female; D005347:Fibroblasts / Q000166:cytology*; D005854:Germ Cells / Q000166:cytology*; D053652:Nuclear Transfer Techniques
D016895:Culture Media, Serum-Free