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PRIMO Vision - Publications
PRIMO Vision is for non-clinical research use only in USA.
Prediction of the in vitro developmental competence of early cleavage stage
mouse embryos with compact timelapse equipment.
Dr. Csaba Pribenszky1,8, Dr. Eszter Losonczi2, Miklós Molnár2, Zsolt Lang3, Szabolcs Mátyás4, Klára Rajczy4, Katalin Molnár4, Dr. Péter Kovács4, Dr. Péter Nagy5, Jason Conceicao6, Dr. Gábor Vajta7
1Department of Animal Breeding and Genetics, Departmet of Animal Breeding and Genetics, Faculty of Veterinary Science, Szent István University, H-1078, István u. 2., Budapest, Hungary
2ARTechnic Co. Research and Development, Csapó u. 13. Debrecen, Hungary
3Department of Biomathematics, Faculty of Veterinary Science, Szent István University, H-1078, István u. 2., Budapest, Hungary
4Department of Embryology, KAÁLI Institute, Budapest, Hungary
5Technical University, Budapest, Hungary
6PIVET Medical Centre, 166-168 Cambridge St, Perth, Western Australia
7Cairns Fertility Centre, Cnr Applin and McLeod St, Cairns, Queensland QLD-4870, Australia
8Correspondence: pribenszky.csaba@aotk.szie.hu
The right preimplantation embryo selection may be improved by a reliable system
capable of early prediction of blastocyst development. Our timelapse investigations
on mouse embryos have proved that the time of the first and second cleavage (to the
2- and 3-cell stages, respectively) has a strong predictive value for further
development in vitro, while cleavage from the 3 cell to the 4 cell stage had no effect
to the blastocyst formation rate. In humans, embryo fragmentations during
preimplantation development have been reported to be associated with lower
pregnancy rates and a higher incidence of developmental abnormalities. Analysis of
our timelapse records has revealed that most fragmentations are reversible in the
mouse, and disappear by resorption in an average of 9 hours. The daily or bi-daily
microscopic checks of embryo development, applied routinely in human IVF
laboratories, would fail to detect 36 or 72% of these fragmentations, respectively.
Fragmentation occurring in a defined time-frame has a strong predictive value to in
vitro embryo development. The compact time-lapse system used in the trial, based
on the 'one camera per patient' principle, has eliminated the usual disadvantages of
timelapse investigations (complicated structure, limited capacity, potential heat or
shear stress, danger of infection) and was found applicable for the routine follow-up
of in vitro embryo development.
Reproductive Biomedicine Online 2009/4551, received 6 Jul 09, sent for refereeing 20 Jul 09, on web 20 Jul 09
The well of the well system: an efficient approach to improve embryo development
G Vajta1,5, T Körösi2, Y Du1, K Nakata3, S Ieda3, M Kuwayama3, ZP Nagy4
1Population Genetics and Embryology, Department of Genetics and Biotechnology, Danish Institute of Agricultural Sciences, Research Centre Foulum, DK-8830 Tjele, Denmark
2Kaáli Intézet, Györ, Hungary
3Advanced Medical Institute of Fertility, Kato Ladies Clinic 7-20-3 Nishishinjuku, Shinjuku-ku, Tokyo, 160-0023, Japan
4Reproductive Biology Associates, 1150 Lake Hearn Drive, Suite 600, Atlanta, GA 30342
5Correspondence: Tel: +45 8999 1262; Fax: =45 8999 1300; e-mail: gabor.vajta@agrsci.dk
Transfer of human embryos at the blastocyst stage may offer considerable benefits including an increased implantation rate and a decreased risk of multiple pregnancies; however, blastocyst culture requires an efficient and reliable in-vitro embryo culture system. In this study, the effect of the well of the well (WOW) system consisting of microwells formed on the bottom of the culture dish was tested in three mammalian species, including humans. The WOW system resulted in significant improvement when comparing the drops for culture of in-vitro-matured and parthenogenetically activated porcine oocytes, and in-vivo-derived mouse zygotes. In human embryos, using a sibling oocyte design, embryos cultured in WOW developed to the blastocyst stage in a significantly higher proportion than did embryos cultured traditionally (55% in WOW and 37% in conventional culture; P < 0.05). In a separate study, also in human, a total of 48 patients with a cumulative 214 unsuccessful previous IVF cycles were selected for the trials. In subsequent intracytoplasmic sperm injection cycles, oocytes/embryos were cultured individually in the WOW system or in microdrops. Transferable quality blastocyst development (48.9% of cultured zygotes) was observed in the WOW system. Ninety-four blastocysts transferred to 45 patients resulted in clinical pregnancy rates of 48.9%, including nine twin pregnancies, seven single pregnancies, five miscarriages and one ectopic pregnancy. The results indicate that the WOW system provides a promising alternative for microdrop culture of mammalian embryos, including human embryos.
Reproductive BioMedicine Online 2008 http://www.rbmonline.com/Article/3282 [e-pub ahead of print on 28 May 2008]
Reproductive BioMedicine Online 2008 Vol. 17 No.1. 73–81
New Method for Culture of Zona-Included or
Zona-Free Embryos: The Well of the Well
(WOW) System
G. VAJTA,1,4* T.T. PEURA,2 P. HOLM,1 A. PA´ LDI,3 T. GREVE,4 A.O. TROUNSON,2 AND H. CALLESEN1
1Embryo Technology Center, Danish Institute of Agricultural Sciences, Research Centre Foulum, Tjele, Denmark
2Centre for Early Human Development, Institute of Reproduction and Development, Monash University,
Clayton, Australia
3Institut Cochin de Ge´ne´tique Mole´culaire, INSERM U 257, Paris, France
4Section of Reproduction, Department of Clinical Studies, Royal Veterinary and Agricultural University,
Frederiksberg C, Denmark
Culture of mammalian zygotes
individually and in small groups results in lower developmental
rates than culture of large groups. Zona-free
zygotes also have impaired developmental potential in
current culture systems. This paper describes a new
approach to resolve the problems, the Well of the Well
(WOW) system. Small wells (WOWs) were formed in
four-well dishes by melting the bottom with heated steel
rods. The WOWs were then rinsed, the wells were filled
with medium, and the embryos were placed into the
WOWs. To test the value of the WOW system a 3 3 3
factorial experiment was performed. Bovine presumptive
zygotes were cultured from day 1 to day 7 (day 0:
day of insemination) using three modules (single embryos,
embryo groups of five, or single zona-digested
embryos) and three different culture systems (400 ml
medium, 200 ml drops, or WOWs). An additional control
group consisted of 40 to 50 embryos cultured in 400 ml
medium. The WOW system resulted in higher blastocyst/oocyte rates for all three modules (single: 59%; group
of five: 61%; single zona-digested: 53%) than the
culture in drops or in wells (P , 0.05 for all). The
developmental rate was independent of the number of
WOWs per well. The cell number of blastocysts cultured
in the WOW system did not differ from that of the
controls. Apart from its theoretical value in revealing the
role of different factors influencing embryo development
in vitro, the WOW system may have immediate
practical consequences in certain areas of mammalian
embryo production.
Mol Reprod Dev 55:256–264,
2000.
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