Triploid human embryonic stem cells derived from tripronuclear zygotes displayed pluripotency and trophoblast differentiation ability similar to the diploid human embryonic stem cells

Abstract

Because the diploid human embryonic stem cells (hESCs) can be successfully derived from tripronuclear zygotes thus, they can serve as an alternative source of derivation of normal karyotype hESC lines. The aim of the present study was to compare the pluripotency and trophoblast differentiation ability of hESCs derived from tripronuclear zygotes and diploid hESCs. In the present study, a total of 20 tripronuclear zygotes were cultured; 8 zygotes developed to the blastocyst stage and 1 hESC line was generated. Unlike the previous studies, chromosomal correction of tripronuclear zygotes during derivation of hESCs did not occur. The established line carries 3 sets of chromosomes and showed a numerical aberration. Although the cell line displayed an abnormal chromosome number, it was found the cell line has been shown to be pluripotent with the ability to differentiate into 3 embryonic germ layers both in vitro and in vivo. The expression of X inactive specific transcript (XIST) in mid-passage (passage 42) of undifferentiated triploid hESCs was detected, indicating X chromosome inactivation of the cell line. Moreover, when this cell line was induced to differentiate toward the trophoblast lineage, morphological and functional trophoblast cells were observed, similar to the diploid hESC line.

Generation, genomic characterization, and differentiation of triploid human embryonic stem cells

“…Overall, the differentiated triploid cells recapitulate many aspects of triploidy-associated pathology, as they displayed a reduced capability of differentiating in vivo and in vitro , and particularly into the neural lineage, suggesting we constructed a reliable model for investigating the effect of triploidy on human development. Triploid hESC lines have been established in the past ( Akyash et al., 2019 ; Baharvand et al., 2006 ; Chen et al., 2012 ; Noggle et al., 2011 ; Rungsiwiwut et al., 2016 ), but they were not characterized extensively. Moreover, in all previous studies, the triploid hESCs differ from their diploid counterparts in their genetic background, in the number of active X chromosomes, and in their imprinting signature.…”

Comparison of Four Protocols for In Vitro Differentiation of Human Embryonic Stem Cells into Trophoblast Lineages by BMP4 and Dual Inhibition of Activin/Nodal and FGF2 Signaling

“…This can be influenced by ambient O 2 conditions, the source and concentration of BMP4 used, starting sizes of colonies, and initial colony density at the time of BMP4 addition [41]. In some reports, FGF2 was present [6,12,16,42] and in other reports, it was excluded at the time BMP4 was added [8,[43][44][45]. For example, Wei and colleagues used a novel E6 medium, which lacks TGF-β and bFGF, to differentiate induced pluripotent stem cells (iPSCs) into trophoblast-like cells.…”

Stem Cell-Based Trophoblast Models to Unravel the Genetic Causes of Human Miscarriages

“…Trophoblast-like cells, derived from BMP4-treated triploid ESC (the product of a tripronuclear zygote), showed morphology, gene expression, and secretion of hCG, similar to a diploid hESC line [ 113 ]. Previously, it has been reported that triploid hESCs have an ability to spontaneously differentiate to the trophoblast lineage, with hCGs detected in the conditioned media at a level of more than 35 mIU/mL [ 114 ].…”