Efficient human sperm pronucleus formation and replication in Xenopus egg extracts.
We have achieved efficient in vitro reactivation and replication of human sperm nuclei in frog egg extracts by constructing a 4-step protocol that mimics the events of fertilization and pronucleus formation in mammalian eggs. With use of this protocol, 78-97% of human sperm nuclei from fertile donors synchronously swelled and completed full genome replication in about 2 h. We document the changes in nuclear structure that accompany efficient DNA synthesis and discuss future research and potential clinical implications of this new system.
Neuber
E
E
Department of Biology, Brandeis University, Waltham, Massachusetts 02454, USA.
Havari
E
E
Aquiles Sanchez
J
J
Powers
R D
RD
Wangh
L J
LJ
D000818:Animals; D002467:Cell Nucleus / Q000502:physiology*; D004261:DNA Replication; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D006801:Humans; D008297:Male; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology*; D014981:Xenopus