Human embryonic stem cells from aneuploid blastocysts identified by pre-implantation genetic screening

Abstract

Human embryonic stem cells are derived from the inner cell mass of pre-implantation embryos. The cells have unlimited proliferation potential and capacity to differentiate into the cells of the three germ layers. Human embryonic stem cells are used to study human embryogenesis and disease modeling and may in the future serve as cells for cell therapy and drug screening. Human embryonic stem cells are usually isolated from surplus normal frozen embryos and were suggested to be isolated from diseased embryos detected by pre-implantation genetic diagnosis. Here we report the isolation of 12 human embryonic stem cell lines and their thorough characterization. The lines were derived from embryos detected to have aneuploidy by pre-implantation genetic screening. Karyotype analysis of these cell lines showed that they are euploid, having 46 chromosomes. Our interpretation is that the euploid cells originated from mosaic embryos, and in vitro selection favored the euploid cells. The undifferentiated cells exhibited long-term proliferation and expressed markers typical for embryonic stem cells such as OCT4, NANOG, and TRA-1-60. The cells manifested pluripotent differentiation both in vivo and in vitro. To further characterize the different lines, we have analyzed their ethnic origin and the family relatedness among them. The above results led us to conclude that the aneuploid mosaic embryos that are destined to be discarded can serve as source for normal euploid human embryonic stem cell lines. These lines represent various ethnic groups; more lines are needed to represent all populations.

TeratoScore: Assessing the Differentiation Potential of Human Pluripotent Stem Cells by Quantitative Expression Analysis of Teratomas

“…Another five were from hPSCs with trisomy of chromosome 12, and four teratomas were initiated from hPSCs with trisomy of chromosome 21. Teratomas were subjected to serial sectioning and H&E staining in order to validate the representation of all three germ layers as previously shown ( Ben-David et al., 2014; Biancotti et al., 2010; Chin et al., 2009; Lavon et al., 2008; Narwani et al., 2010 ). In many histological analyses, structures from all three embryonic germ layers were found in the same section, supporting the notion that different cell types exist in close proximity in the teratomas ( Figure S1 ).…”

Aneuploidy induces profound changes in gene expression, proliferation and tumorigenicity of human pluripotent stem cells

“…CSES 18,24 and HUES 25 hESCs and hiPS18 hiPSCs 26 were cultured under standard culture conditions, as previously described 7 . In short, cells were grown on mitomycin C-treated MEF feeder layer (obtained from 13.5-day embryos) in 85% KnockOut Dulbecco's modified Eagle's medium (Invitrogen) supplemented with 15% KnockOut SR (a serum-free formulation; Invitrogen), 1 mM glutamine, 0.1 mM b-mercaptoethanol (Sigma-Aldrich), 1% nonessential amino acids stock (Invitrogen), penicillin (50 U ml À 1 ), streptomycin (50 mg ml À 1 ) and FGF2 (Invitrogen).…”

Analysis of chromosomal aberrations and recombination by allelic bias in RNA-Seq

“…Human ESCs (CSES 37 38 and HUES14 (ref. 39 )) were cultured on mouse embryonic fibroblast treatment with mitomycin-C. Culture medium contained KnockOut Dulbecco's modified Eagle's medium (Gibco-Invitrogen, CA) supplemented with 15% KnockOut-SR (Gibco-Invitrogen, CA), 1 mM glutamine, 0.1 mM β-mercaptoethanol (Sigma-Aldrich, MO), 1% non-essential amino-acid stock (Gibco-Invitrogen, CA), penicillin (50 U ml −1 ), streptomycin (50 μg ml −1 ), and 8 ng ml −1 fibroblast growth factor 2 (Gibco-Invitrogen, CA).…”