title: Generation of progeny from embryonic stem cells by microinsemination of male germ cells from chimeric mice.
abstract: Mice chimeric for embryonic stem (ES) cells have not always successfully produced ES-derived offspring. Here we show that the male gametes from ES cells could be selected in male chimeric mice testes by labeling donor ES cells or host blastocytes with GFP. Male GFP-expressing ES-derived germ cells occurred as colonies in the chimeric testes, where the seminiferous tubules were separated into green and non-green regions. When mature spermatozoa from green tubules were used for microinsemination, GFP-expressing offspring were efficiently obtained. Using a reverse study, we also obtained ES-derived progeny from GFP-negative ES cells in GFP-labeled host chimeras. Furthermore, we showed this approach could be accelerated by using round spermatids from the testes of 20-day-old chimeric mice. Thus, this technique allowed us to generate the ES cell-derived progeny even from the low contributed chimeric mice, which cannot produce ES-origin offspring by natural mating.
authors.affiliation: Laboratory for Genomic Reprogramming, Center for Developmental Biology, Riken Kobe, Japan. mizutani@cdb.riken.jp
authors.forename: Eiji
authors.initials: E
authors.lastname: Mizutani
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ohta
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kishigami
authors.forename: Nguyen
authors.initials: N
authors.lastname: Van Thuan
authors.forename: Takafusa
authors.initials: T
authors.lastname: Hikichi
authors.forename: Sayaka
authors.initials: S
authors.lastname: Wakayama
authors.forename: Eimei
authors.initials: E
authors.lastname: Sato
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Wakayama
mesh_terms: D000818:Animals; D002460:Cell Line; D002678:Chimera / Q000235:genetics*; D019976:Cloning, Organism; D005260:Female; D017930:Genes, Reporter; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D008845:Microinjections; D013087:Spermatids / Q000637:transplantation; D033581:Stem Cell Transplantation / Q000379:methods; D013737:Testis / Q000166:cytology