Production of cloned pigs from adult somatic cells by chemically assisted removal of maternal chromosomes.
The present study demonstrated that brief treatment of in vitro-matured porcine oocytes with demecolcine results in a membrane protrusion that contains a condensed chromosome mass, which can be easily removed by aspiration. This simple, chemically assisted method for removing maternal chromosomes enabled the production of a large number of nuclear-transferred porcine eggs. The development of eggs whose chromosomes were removed by this procedure following transfer of somatic cell nuclei to the blastocyst stage was not significantly different among groups activated using different procedures (6% to 11%) and was also not different among donor cells of different origins (3% to 9%), except for cumulus cells (0.4%). After transfer of 180 to 341 nuclear-transferred eggs that received somatic cells to 6 recipients, 2 of the recipients produced 8 healthy cloned piglets from the heart cells of a female pig. The chemically assisted method for removing maternal chromosomes was also effective for bovine and rabbit eggs.
Yin
Xi Jun
XJ
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, 3327-204 Nakamachi, Nara 631-8505, Japan.
Tani
Tetsuya
T
Yonemura
Isao
I
Kawakami
Masahiro
M
Miyamoto
Kazunori
K
Hasegawa
Rie
R
Kato
Yoko
Y
Tsunoda
Yukio
Y
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D002417:Cattle; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D002875:Chromosomes / Q000187:drug effects / Q000235:genetics* / Q000648:ultrastructure; D019976:Cloning, Organism / Q000379:methods*; D003703:Demecolcine / Q000494:pharmacology; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D018895:Microsatellite Repeats; D009206:Myocardium / Q000166:cytology; D011247:Pregnancy; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine
D000972:Antineoplastic Agents, Phytogenic; D003703:Demecolcine