title: Derivation of normal diploid human embryonic stem cells from tripronuclear zygotes with analysis of their copy number variation and loss of heterozygosity.
abstract: This study sought to establish archives of genetic copy number variation (CNV) in human embryonic stem cell (hESC) lines that are associated with known diseases. We collected patients' fresh, discarded zygotes from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) protocols. A total of 208 fresh, tripronuclear, discarded zygotes were also collected in this study from patients on the third day of their treatment cycle, prior to transfer. The blastula-formation rates were 13.51% (26/192) and 26.7% (4/15) while the high-quality blastocyst formation rates were 5.8% (11/192) and 20% (3/15) in the IVF and ICSI groups, respectively. The inner cell mass (ICM) from each embryo was mechanically separated, and then grown on feeder layers consisting of mouse embryonic fibroblasts and human foreskin fibroblasts (a 1:1 mixture). The hESC karyotype was determined by traditional G-banding; analysis of the results for the Zh19P25 and Zh20P24 cell lines showed that both were 46 XY. CNV and loss-of-heterozygosity analysis of hESC gDNA was performed to assess the genetic characteristics associated with molecular diseases using the high-resolution Infinium High-Density HumanCytoSNP-12 DNA chip. Seven CNVs in Zh19P25 and Zh20P24 were deletions, and a region that corresponds to Potocki-Shaffer disease, 11p11.2-11p11.12 in Zh20P24, showed a 2.98-Mb loss. These data together suggest that single-nucleotide polymorphism (SNP) microarray analysis for molecular cytogenetic features can help to distinguish hESC lines with a normal karyotype from tripronuclear zygotes with known, disease-related characteristics.
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.|Department of Human Anatomy, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Chen
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Wenbin
authors.initials: W
authors.lastname: Niu
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Wenzhu
authors.initials: W
authors.lastname: Yu
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Shanjun
authors.initials: S
authors.lastname: Dai
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Kong
authors.affiliation: Department of Obstetrics and Gynecology, Stanford University Medical Center, Palo Alto, California.
authors.forename: Yimin
authors.initials: Y
authors.lastname: Shu
authors.affiliation: Reproductive Medical Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
authors.forename: Yingpu
authors.initials: Y
authors.lastname: Sun
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D056915:DNA Copy Number Variations; D004171:Diploidy; D046149:Embryo Culture Techniques / Q000379:methods; D005307:Fertilization in Vitro; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007621:Karyotyping; D019656:Loss of Heterozygosity; D051379:Mice; D020554:Sperm Injections, Intracytoplasmic; D015053:Zygote / Q000166:cytology* / Q000378:metabolism