Generation of medaka fish haploid embryonic stem cells.
Haploid embryonic stem (ES) cells combine haploidy and pluripotency, enabling direct genetic analyses of recessive phenotypes in vertebrate cells. Haploid cells have been elusive for culture, due to their inferior growth and genomic instability. Here, we generated gynogenetic medaka embryos and obtained three haploid ES cell lines that retained pluripotency and competitive growth. Upon nuclear transfer into unfertilized oocytes, the haploid ES cells, even after genetic engineering, generated viable offspring capable of germline transmission. Hence, haploid medaka ES cells stably maintain normal growth, pluripotency, and genomic integrity. Mosaic oocytes created by combining a mitotic nucleus and a meiotic nucleus can generate fertile fish offspring. Haploid ES cells may offer a yeast-like system for analyzing recessive phenotypes in numerous cell lineages of vertebrates in vitro.
Yi
Meisheng
M
Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260.
Hong
Ni
N
Hong
Yunhan
Y
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape; D043171:Chromosomal Instability; D019976:Cloning, Organism; D003433:Crosses, Genetic; D004171:Diploidy; D004625:Embryo, Nonmammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005260:Female; D006238:Haploidy; D008297:Male; D053652:Nuclear Transfer Techniques; D009865:Oocytes; D009990:Oryzias / Q000196:embryology / Q000235:genetics / Q000502:physiology; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D018183:Transplantation Chimera