Pronuclear Removal of Tripronuclear Zygotes Can Establish Heteroparental Normal Karyotypic Human Embryonic Stem Cells

Overview

Journal J Assist Reprod Genet

Publisher Springer

Specialties Genetics
Reproductive Medicine

Date 2016 Jan 6

PMID 26727933

Citations 2

Authors

Hong-Qing Liao

Qi Ouyang

Shuo-Ping Zhang

De-Hua Cheng

Guang-Xiu Lu

Ge Lin

Affiliations

Soon will be listed here.

Abstract

Purpose: This study aimed to derive heteroparental normal karyotypic human embryonic stem cells (hESCs) from microsurgically corrected tripronuclear (3PN) zygotes.

Methods: After sequential culture for 5-6 days, embryos developed from microsurgically corrected 3PN zygotes were analyzed by fluorescence in situ hybridization (FISH) using probes for chromosomes 17, X and Y. Intact 3PN zygotes from clinical in vitro fertilization (IVF) cycles were cultured as the control group. The inner cell mass (ICM) of blastocysts that developed from microsurgically corrected 3PN zygotes was used to derive hESC lines, and the stem cell characteristics of these lines were evaluated. G-banding analysis was adopted to identify the karyotype of the hESC line, and the heteroparental inheritance of the hESC line was analyzed by DNA fingerprinting analysis.

Results: The blastocyst formation rate (13.5 %) of the microsurgically corrected 3PN zygotes was significantly higher (P < 0.05) than that of intact 3PN zygotes (8.7 %). The diploid rate of the blastocysts (55.0 %) was significantly higher (P < 0.05) than that of the arrested cleavage-stage embryos (18.4 %) in microsurgically corrected 3PN zygotes. The triploid rate of the microsurgically corrected 3PN zygotes (5.7 %) was significantly lower (P < 0.01) than that of intact 3PN zygotes (19.4 %). Furthermore, we established one heteroparental normal karyotypic hESC line from the microsurgically corrected tripronuclear zygotes.

Conclusions: Pronuclear removal can effectively remove the surplus chromosome set of 3PN zygotes. A combination of pronuclear removal and blastocyst culture enables the selection of diploidized blastocysts from which heteroparental normal karyotypic hESC lines can be derived.

Citing Articles

Histone demethylase KDM4A overexpression improved the efficiency of corrected human tripronuclear zygote development.

Zhu H, Kang X, Jin L, Zhang P, Wu H, Tan T Mol Hum Reprod. 2021; 27(3).

PMID: 33599278 PMC: 7939728. DOI: 10.1093/molehr/gaab012.

Time-lapse observation and transcriptome analysis of a case with repeated multiple pronuclei after IVF/ICSI.

Dai J, Leng L, Lu C, Gong F, Zhang S, Zheng W J Assist Reprod Genet. 2017; 34(9):1189-1197.

PMID: 28643089 PMC: 5581788. DOI: 10.1007/s10815-017-0972-9.

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