nach oben

Journal of Assisted Reproduction and Genetics

Erschienen in:

01.02.2016 | Stem Cell Biology

Pronuclear removal of tripronuclear zygotes can establish heteroparental normal karyotypic human embryonic stem cells

verfasst von: Hong-Qing Liao, Qi OuYang, Shuo-Ping Zhang, De-Hua Cheng, Guang-Xiu Lu, Ge Lin

Erschienen in: Journal of Assisted Reproduction and Genetics | Ausgabe 2/2016

Einloggen, um Zugang zu erhalten

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.

Pieters MH, Dumoulin JC, Ignoul-Vanvuchelen RC, Bras M, Evers JL, Geraedts JP. Triploidy after in vitro fertilization: cytogenetic analysis of human zygotes and embryos. J Assist Reprod Genet. 1992;9:68–76.CrossRefPubMed

Rosenbusch BE. Selective microsurgical removal of a pronucleus from tripronuclear human oocytes to restore diploidy: disregarded but valuable? Fertil Steril. 2009;92:897–903. doi:10.1016/j.fertnstert.2008.07.1740.CrossRefPubMed

Porter R, Han T, Tucker MJ, Graham J, Liebermann J, Sills ES. Estimation of second polar body retention rate after conventional insemination and intracytoplasmic sperm injection: in vitro observations from more than 5000 human oocytes. J Assist Reprod Genet. 2003;20:371–6.PubMedCentralCrossRefPubMed

Kang HJ, Rosenwaks Z. Triploidy—the breakdown of monogamy between sperm and egg. Int J Dev Biol. 2008;52:449–54. doi:10.1387/ijdb.082602hk.CrossRefPubMed

Plachot M, Crozet N. Fertilization abnormalities in human in-vitro fertilization. Hum Reprod. 1992;7 Suppl 1:89–94.CrossRefPubMed

Staessen C, Van Steirteghem AC. The chromosomal constitution of embryos developing from abnormally fertilized oocytes after intracytoplasmic sperm injection and conventional in-vitro fertilization. Hum Reprod. 1997;12:321–7.CrossRefPubMed

Grossmann M, Calafell JM, Brandy N, Vanrell JA, Rubio C, Pellicer A, et al. Origin of tripronucleate zygotes after intracytoplasmic sperm injection. Hum Reprod. 1997;12:2762–5.CrossRefPubMed

Golubovsky MD. Postzygotic diploidization of triploids as a source of unusual cases of mosaicism, chimerism and twinning. Hum Reprod. 2003;18:236–42.CrossRefPubMed

Rosenbusch BE. Mechanisms giving rise to triploid zygotes during assisted reproduction. Fertil Steril. 2008;90:49–55. doi:10.1016/j.fertnstert.2007.06.031.CrossRefPubMed

10.

Zaragoza MV, Surti U, Redline RW, Millie E, Chakravarti A, Hassold TJ. Parental origin and phenotype of triploidy in spontaneous abortions: predominance of diandry and association with the partial hydatidiform mole. Am J Hum Genet. 2000;66:1807–20. doi:10.1086/302951.PubMedCentralCrossRefPubMed

11.

McFadden DE, Robinson WP. Phenotype of triploid embryos. J Med Genet. 2006;43:609–12. doi:10.1136/jmg.2005.037747.PubMedCentralCrossRefPubMed

12.

Berkowitz RS, Goldstein DP. Clinical practice molar pregnancy. N Engl J Med. 2009;360:1639–45. doi:10.1056/NEJMcp0900696.CrossRefPubMed

13.

Rawlins RG, Binor Z, Radwanska E, Dmowski WP. Microsurgical enucleation of tripronuclear human zygotes. Fertil Steril. 1988;50:266–72.PubMed

14.

Gordon JW, Grunfeld L, Garrisi GJ, Navot D, Laufer N. Successful microsurgical removal of a pronucleus from tripronuclear human zygotes. Fertil Steril. 1989;52:367–72.PubMed

15.

Malter HE, Cohen J. Embryonic development after microsurgical repair of polyspermic human zygotes. Fertil Steril. 1989;52:373–80.PubMed

16.

Palermo G, Munne S, Cohen J. The human zygote inherits its mitotic potential from the male gamete. Hum Reprod. 1994;9:1220–5.PubMed

17.

Ivakhnenko V, Cieslak J, Verlinsky Y. A microsurgical technique for enucleation of multipronuclear human zygotes. Hum Reprod. 2000;15:911–6.CrossRefPubMed

18.

Escriba MJ, Martin J, Rubio C, Valbuena D, Remohi J, Pellicer A, et al. Heteroparental blastocyst production from microsurgically corrected tripronucleated human embryos. Fertil Steril. 2006;86:1601–7. doi:10.1016/j.fertnstert.2006.04.047.CrossRefPubMed

19.

Gu Y-F, Lin G, Lu C-F, Lu G-X. Analysis of the first mitotic spindles in human in vitro fertilized tripronuclear zygotes after pronuclear removal. Reprod BioMed Online. 2009;19:745–54.CrossRefPubMed

20.

Kattera S, Chen C. Normal birth after microsurgical enucleation of tripronuclear human zygotes: case report. Hum Reprod. 2003;18:1319–22.CrossRefPubMed

21.

Gardner DK, Schoolcraft WB. No longer neglected: the human blastocyst. Hum Reprod. 1998;13:3289–92.CrossRefPubMed

22.

Jones GM, Trounson AO, Gardner DK, Kausche A, Lolatgis N, Wood C. Evolution of a culture protocol for successful blastocyst development and pregnancy. Hum Reprod. 1998;13:169–77.CrossRefPubMed

23.

Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998;282:1145–7.CrossRefPubMed

24.

Reubinoff BE, Pera MF, Fong CY, Trounson A, Bongso A. Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol. 2000;18:399–404. doi:10.1038/74447.CrossRefPubMed

25.

Mitalipova M, Calhoun J, Shin S, Wininger D, Schulz T, Noggle S, et al. Human embryonic stem cell lines derived from discarded embryos. Stem Cells. 2003;21:521–6. doi:10.1634/stemcells.21-5-521.CrossRefPubMed

26.

Lavon N, Narwani K, Golan-Lev T, Buehler N, Hill D, Benvenisty N. Derivation of euploid human embryonic stem cells from aneuploid embryos. Stem Cells. 2008;26:1874–82. doi:10.1634/stemcells.2008-0156.CrossRefPubMed

27.

Lin G, OuYang Q, Zhou XY, Gu YF, Yuan D, Li W, et al. A highly homozygous and parthenogenetic human embryonic stem cell line derived from a one-pronuclear oocyte following in vitro fertilization procedure. Cell Res. 2007;17:999–1007. doi:10.1038/cr.2007.97.CrossRefPubMed

28.

Lin G, Xie Y, OuYang Q, Qian X, Xie P, Zhou X, et al. HLA-matching potential of an established human embryonic stem cell bank in China. Cell Stem Cell. 2009;5:461–5. doi:10.1016/j.stem.2009.10.009.CrossRefPubMed

29.

Angell RR, Templeton AA, Messinis IE. Consequences of polyspermy in man. Cytogenet Cell Genet. 1986;42:1–7.CrossRefPubMed

30.

Kola I, Trounson A, Dawson G, Rogers P. Tripronuclear human oocytes: altered cleavage patterns and subsequent karyotypic analysis of embryos. Biol Reprod. 1987;37:395–401.CrossRefPubMed

31.

Rosenbusch B, Schneider M, Sterzik K. The chromosomal constitution of multipronuclear zygotes resulting from in-vitro fertilization. Hum Reprod. 1997;12:2257–62.CrossRefPubMed

32.

Iliopoulos D, Vassiliou G, Sekerli E, Sidiropoulou V, Tsiga A, Dimopoulou D, et al. Long survival in a 69, XXX triploid infant in Greece. Genet Mol Res. 2005;4:755–9.PubMed

33.

Clouston HJ, Fenwick J, Webb AL, Herbert M, Murdoch A, Wolstenholme J. Detection of mosaic and non-mosaic chromosome abnormalities in 6- to 8-day old human blastocysts. Hum Genet. 1997;101:30–6.CrossRefPubMed

34.

Evsikov S, Verlinsky Y. Mosaicism in the inner cell mass of human blastocysts. Hum Reprod. 1998;13:3151–5.CrossRefPubMed

35.

Magli MC, Jones GM, Gras L, Gianaroli L, Korman I, Trounson AO. Chromosome mosaicism in day 3 aneuploid embryos that develop to morphologically normal blastocysts in vitro. Hum Reprod. 2000;15:1781–6.CrossRefPubMed

36.

Sandalinas M, Sadowy S, Alikani M, Calderon G, Cohen J, Munne S. Developmental ability of chromosomally abnormal human embryos to develop to the blastocyst stage. Hum Reprod. 2001;16:1954–8.CrossRefPubMed

37.

Liao H, Zhang S, Cheng D, Ouyang Q, Lin G, Gu Y, et al. Cytogenetic analysis of human embryos and embryonic stem cells derived from monopronuclear zygotes. J Assist Reprod Genet. 2009;26:583–9. doi:10.1007/s10815-009-9355-1.PubMedCentralCrossRefPubMed

38.

Hardy K. Cell death in the mammalian blastocyst. Mol Hum Reprod. 1997;3:919–25.CrossRefPubMed

39.

Janny L, Menezo YJ. Maternal age effect on early human embryonic development and blastocyst formation. Mol Reprod Dev. 1996;45:31–7.CrossRefPubMed

40.

Fan Y, Li R, Huang J, Yu Y, Qiao J. Diploid, but not haploid, human embryonic stem cells can be derived from microsurgically repaired tripronuclear human zygotes. Cell Cycle. 2013;12:302–11. doi:10.4161/cc.23103.PubMedCentralCrossRefPubMed

41.

Jiang C, Cai L, Huang B, Dong J, Chen A, Ning S, et al. Normal human embryonic stem cell lines were derived from microsurgical enucleated tripronuclear zygotes. J Cell Biochem. 2013;114:2016–23. doi:10.1002/jcb.24547.CrossRefPubMed

42.

Fan Y, Li R, Huang J, Zhao HC, Ding T, Sun X, et al. Improved efficiency of microsurgical enucleated tripronuclear zygotes development and embryonic stem cell derivation by supplementing epidermal growth factor, brain-derived neurotrophic factor, and insulin-like growth factor-1. Stem Cells Dev. 2014;23:563–75. doi:10.1089/scd.2013.0420.PubMedCentralCrossRefPubMed

Titel: Pronuclear removal of tripronuclear zygotes can establish heteroparental normal karyotypic human embryonic stem cells
verfasst von: Hong-Qing Liao
Qi OuYang
Shuo-Ping Zhang
De-Hua Cheng
Guang-Xiu Lu
Ge Lin
Publikationsdatum: 01.02.2016
Verlag: Springer US
Erschienen in: Journal of Assisted Reproduction and Genetics / Ausgabe 2/2016
Print ISSN: 1058-0468
Elektronische ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-015-0634-8

Neu im Fachgebiet Gynäkologie und Geburtshilfe

Mammakarzinom: Senken Statine das krebsbedingte Sterberisiko?

15.05.2024 Mammakarzinom Nachrichten

Frauen mit lokalem oder metastasiertem Brustkrebs, die Statine einnehmen, haben eine niedrigere krebsspezifische Mortalität als Patientinnen, die dies nicht tun, legen neue Daten aus den USA nahe.

Neue S3-Leitlinie zur unkomplizierten Zystitis: Auf Antibiotika verzichten?

15.05.2024 Harnwegsinfektionen Nachrichten

Welche Antibiotika darf man bei unkomplizierter Zystitis verwenden und wovon sollte man die Finger lassen? Welche pflanzlichen Präparate können helfen? Was taugt der zugelassene Impfstoff? Antworten vom Koordinator der frisch überarbeiteten S3-Leitlinie, Prof. Florian Wagenlehner.

Gestationsdiabetes: In der zweiten Schwangerschaft folgenreicher als in der ersten

13.05.2024 Gestationsdiabetes Nachrichten

Das Risiko, nach einem Gestationsdiabetes einen Typ-2-Diabetes zu entwickeln, hängt nicht nur von der Zahl, sondern auch von der Reihenfolge der betroffenen Schwangerschaften ab.

Embryotransfer erhöht womöglich Leukämierisiko der Kinder

13.05.2024 Assistierte Reproduktion Nachrichten

Reproduktionsmedizinische Techniken haben theoretisch das Potenzial, den epigenetischen Code zu verändern und somit das Krebsrisiko der Kinder zu erhöhen. Zwischen Embryotransfer und Leukämie scheint sich ein solcher Zusammenhang bestätigt zu haben.

Update Gynäkologie

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert – ganz bequem per eMail.

Newsletter bestellen

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusions

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 2/2016

Impact of sperm DNA chromatin in the clinic

Heads or tails: cell biological secrets of the spermatozoon revealed

Agonist depot versus OCP programming of frozen embryo transfer: a retrospective analysis of freeze-all cycles

Morphokinetics of embryos developed from oocytes matured in vitro