Comparative evaluation of human embryonic stem cell lines derived from zygotes with normal and abnormal pronuclei

Qin, Huan; Gao, Xuan; Wang, Yan; Shen, Yun; Ma, Wu; Chen, Zi‐Jiang

doi:10.1002/dvdy.22175

Cited by 17 publications

(29 citation statements)

References 22 publications

(25 reference statements)

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“…The data suggested the following: (1) the diploid hESC lines originated from the tripronucear zygotes that arose from the third pattern of division; and (2) the in vitro selection favored diploid cells. This is in accordance with previously reported results [11,12,29]. The present report is, we believe, the first record of deriving normal and abnormal hESC lines from 3PN zygotes and comparing the characteristics of these two kinds of hESC lines.…”

Section: In Vitro and In Vivo Differentiation Potential Of The Hesc Linesupporting

confidence: 94%

“…Our results suggested that in the modified culture medium the developmental potential of the 3PN zygotes was lower than that of control group, but the efficiency of the hESC derivation was similar in the two group. According to the data in those publications [10][11][12], there is also no difference in the derivation efficiency of hESCs from mononuclear and tripronuclear zygotes between our lab and other labs ( Table 2).…”

Section: In Vitro and In Vivo Differentiation Potential Of The Hesc Linesupporting

confidence: 50%

“…According to the data in those publications [10][11][12], there is no difference in the derivation efficiency of hESCs from abnormal fertilized zygotes between our lab and other labs ( Table 2). …”

Section: Resultsmentioning

confidence: 55%

“…In our IVF lab,~50% of the couples having abnormal fertilized zygotes agreed to donate them for derivation of hESC lines, a percentage that is much higher than the percentage (~20%) of couples agreeing to donate euploid spare embryos. A few groups have derived diploid hESC lines from both a mononuclear zygote [10] and tripronuclear zygotes [11,12]. However, it is necessary to investigate the efficiency and the ratio of deriving abnormal and normal hESC lines from tripronuclear zygotes to know more about the developmental potential of abnormal zygotes.…”

Section: In Vitro and In Vivo Differentiation Potential Of The Hesc Linementioning

confidence: 99%

“…Recently, several groups have reported that diploid hESC lines were derived from a mononuclear zygote [10] and tripronuclear zygotes [11,12]. However, no earlier deriving abnormal and normal hESC line at the same time has been reported from tripronuclear zygotes.…”

Section: Introductionmentioning

confidence: 99%

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Triploid and diploid embryonic stem cell lines derived from tripronuclear human zygotes

Chen

Luo

Fan

et al. 2012

J Assist Reprod Genet

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Purpose Human embryonic stem cells (hESCs) are selfrenewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. The need for new hESC lines motivated our attempts to find a new resource for the derivation of hESC lines. The aim of this work was to establish more hESC lines from abnormal fertilized zygotes and to meet the emerging requirements for their use in cell replacement therapies, disease modeling, and basic research. Methods A total of 130 tripronuclear human zygotes was collected 18-20 h post-insemination and cultured in a modified culture medium. The inner cell mass of 12 blastocysts were isolated by a mechanical method in order to establish embryonic stem cell lines. Results We established four hESC lines derived from 130 trinuclear zygotes, one of which was triploid and the others were diploid. The efficiency of deriving hESC lines is 3.08%. The ratio of deriving triploid and diploid hESC lines is 1:3. All of these hESC lines exhibited similar markers of undifferentiated hESCs and had the typical morphology of hESCs, a capacity for long-term proliferation, and pluripotent differentiation potential both in vivo and in vitro.Conclusions These abnormal zygotes, which otherwise would have been discarded, can serve as an alternative source for normal euploid hESC lines.

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Section: In Vitro and In Vivo Differentiation Potential Of The Hesc Linesupporting

confidence: 94%

Section: In Vitro and In Vivo Differentiation Potential Of The Hesc Linesupporting

confidence: 50%

Section: Resultsmentioning

confidence: 55%

Section: In Vitro and In Vivo Differentiation Potential Of The Hesc Linementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Triploid and diploid embryonic stem cell lines derived from tripronuclear human zygotes

Chen

Luo

Fan

et al. 2012

J Assist Reprod Genet

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Normal human embryonic stem cell lines were derived from microsurgical enucleated tripronuclear zygotes

Jiang

Cai

Huang

et al. 2013

J of Cellular Biochemistry

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A normal fertilized human zygote contains two pronuclei, but zygotes may also display one, three, or even more pronuclei resulting from irregular insemination or meiotic division. Today diploid and triploid human embryonic stem cell (hESC) lines have been derived from tripronuclear (3PN) triploid zygotes, and an in-vitro fertilization (IVF) baby was born from a rescued diploid zygote by removing the extra male pronucleus of the 3PN zygote. However, whether hESCs can be derived from a rescued 3PN zygote is still unknown. Here, by microsurgical pronuclear removal, we restored 61 diploid zygotes from 3PN zygotes donated by 35 couples, and 11 blastocysts developed with a blastocyst rate of 18.0%, which seems higher than that of nonrescued 3PN zygotes according to previous reports. After the whole zona pellucida free embryos were plated onto feeder cells to grow and passage, 2 hESC lines (CCRM-hESC-22 and CCRM-hESC-23) were generated and both carried normal karyotype (46, XY). The hESC lines were then characterized by morphology, expansion in vitro, and expression of specific markers of alkaline phosphatase, OCT4, SSEA4, TRA-1-60 and TRA-1-81. Furthermore, the pluripotency of these 2 hESC lines was confirmed by in vitro embryoid body formation and in vivo teratoma production. Our study indicates that depronucleared 3PN zygotes can improve the blastocysts formation rate, and normal hESC lines can be derived from those corrected 2PN embryos. Based on their multi-directional differentiation potential in vitro, the established hESC lines could be applied to the developmental risk assessment for IVF babies born from restored zygotes.

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Abnormalities in centrosome number in human embryos and embryonic stem cells

Ouyang

Dai

et al. 2016

Molecular Reproduction Devel

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Chromosomal abnormalities are common in human embryos. Previous studies have suggested links between centrosome number and chromosome abnormalities, but information regarding abnormalities in centrosome number in human embryos is limited. We analyzed abnormalities in centrosome number in human embryos and embryonic stem cells (hESCs). Following normal fertilization, supernumerary centrosomes were present at rates of 7.3% in two-pronucleus (2PN)-stage zygotes and 6.5% in first-cleavage zygotes. Supernumerary centrosomes were also detected in 24.4% of blastomeres from 60% of embryos derived from 2PN zygotes. Conversely, in mono- (1PN) and tri-pronucleus (3PN) zygotes, the frequency of abnormal centrosome number increased substantially at first cleavage. Rates in blastomeres of Day-3 embryos, however, were about the same between embryos derived from 1PN and 2PN zygotes, whereas abnormalities in centrosome number were higher in those from 3PN zygotes. By comparison, the rate of abnormal centrosome numbers in hESCs was 1.5-11.2%. Thus, abnormalities in centrosome number existed in human zygotes and cleaved embryos-especially those resulting from aberrant fertilization-but the frequency of such abnormalities was lower in hESCs derived from these embryos. These findings identify a source of the chromosomal instability in human embryos and hESCs, and highlight new safety issues for human assisted reproductive technology. Mol. Reprod. Dev. 83: 392-404, 2016. © 2016 Wiley Periodicals, Inc.

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Comparative evaluation of human embryonic stem cell lines derived from zygotes with normal and abnormal pronuclei

Cited by 17 publications

References 22 publications

Triploid and diploid embryonic stem cell lines derived from tripronuclear human zygotes

Triploid and diploid embryonic stem cell lines derived from tripronuclear human zygotes

Normal human embryonic stem cell lines were derived from microsurgical enucleated tripronuclear zygotes

Abnormalities in centrosome number in human embryos and embryonic stem cells

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