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| Production of early monozygotic twin bovine embryos in vitro by the blastomere separation and coculture technique |
| ZHAO Shan-jiang, ZHAO Xue-ming, DU Wei-hua, HAO Hai-sheng, LIU Yan, QIN Tong, WANG Dong |
| Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China |
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摘要 The objective of this study was to establish an efficient system of producing early monozygotic twin bovine embryos in vitro using the blastomere separation and coculture technique. In this study, early eight-cell embryos were chosen to optimize the separation method, and multi-coculture tactics were applied to improve the efficiency of this production system. Bovine embryo blastomeres (groups of at least 30 at the eight-cell stage) were separated into eight segments (to regard an eight-cell embryo as a tangerine, a blastomere as one segment) and one, two and four segments (blastomeres) were cultured respectively in microwells on the bottom of the four-well dish (Nunc, Denmark) with 400 μL of culture medium under paraffin oil. Four different types of coculture tactics (cocultured with nothing, intact embryos, bovine cumulus cells (bCCs), intact embryos & bCCs) were applied to the group of four segments (blastomeres). Finally, diameter and inner cell mass (ICM):trophectoderm (TE) cell ratio was measured as a criterion to assess the quality of the twin embryos which were derived from bovine separated blastomeres. Our results showed that rate of blastocyst formation of the four segments group was significantly greater than one or two group (P<0.05). In addition, rate of blastocyst formation was significantly increased when the four segments were cocultured with intact embryo & bCCs (P<0.05). Although the ICM, TE and total cells of blastocysts derived from separated blastomeres was less than the control group from intact embryo (P<0.05), more important quality indicator of the blastocyst diameter and ICM:TE cell ratio was similar between our experimental group and the control group (P>0.05). Thus, these results suggest that combined with intact embryos & bCCs coculture system, culturing four isolated segments (blastomeres) per microwell is an efficient system of producing early monozygotic twin bovine embryos. Furthermore, our results also indicate that the quality of blastocysts derived from separated blastomere may be similar to those derived from intact eight-cell embryos.
Abstract The objective of this study was to establish an efficient system of producing early monozygotic twin bovine embryos in vitro using the blastomere separation and coculture technique. In this study, early eight-cell embryos were chosen to optimize the separation method, and multi-coculture tactics were applied to improve the efficiency of this production system. Bovine embryo blastomeres (groups of at least 30 at the eight-cell stage) were separated into eight segments (to regard an eight-cell embryo as a tangerine, a blastomere as one segment) and one, two and four segments (blastomeres) were cultured respectively in microwells on the bottom of the four-well dish (Nunc, Denmark) with 400 μL of culture medium under paraffin oil. Four different types of coculture tactics (cocultured with nothing, intact embryos, bovine cumulus cells (bCCs), intact embryos & bCCs) were applied to the group of four segments (blastomeres). Finally, diameter and inner cell mass (ICM):trophectoderm (TE) cell ratio was measured as a criterion to assess the quality of the twin embryos which were derived from bovine separated blastomeres. Our results showed that rate of blastocyst formation of the four segments group was significantly greater than one or two group (P<0.05). In addition, rate of blastocyst formation was significantly increased when the four segments were cocultured with intact embryo & bCCs (P<0.05). Although the ICM, TE and total cells of blastocysts derived from separated blastomeres was less than the control group from intact embryo (P<0.05), more important quality indicator of the blastocyst diameter and ICM:TE cell ratio was similar between our experimental group and the control group (P>0.05). Thus, these results suggest that combined with intact embryos & bCCs coculture system, culturing four isolated segments (blastomeres) per microwell is an efficient system of producing early monozygotic twin bovine embryos. Furthermore, our results also indicate that the quality of blastocysts derived from separated blastomere may be similar to those derived from intact eight-cell embryos.
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Received: 11 December 2014
Accepted:
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| Fund: The work was supported by the China Agriculture Research System (CARS-37) and the Agricultural Science and Technology Innovation Program, China (ASTIP-IAS06-2015). |
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Corresponding Authors:
ZHU Hua-bin,Tel: +86-10-62815892, Fax: +86-10-62895971,E-mail: zhuhuabin@caas.cn
E-mail: zhuhuabin@caas.cn
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| About author: ZHAO Shan-jiang, Mobile: +86-18612246578,E-mail: shanjiangzhao@126.com; |
Cite this article:
ZHAO Shan-jiang, ZHAO Xue-ming, DU Wei-hua, HAO Hai-sheng, LIU Yan, QIN Tong, WANG Dong.
2015.
Production of early monozygotic twin bovine embryos in vitro by the blastomere separation and coculture technique. Journal of Integrative Agriculture, 14(10): 2034-2041.
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| Allen W R, Pashen R L. 1984. Production of monozygotic (identical) horse twins by embryo micromanipulation.Journal of Reproduction and Fertility, 71, 607-613Brackett B G, Oliphant G. 1975. Capacitation of rabbitspermatozoa in vitro. Biology of Reproduction, 12, 260-274Dang-Nguyen T Q, Kaneda M, Somfai T, Haraguchi S,Matsukawa K, Akagi S, Kikuchi K, Nakai M, Nguyen B X,Tajima A, Kanai Y, Nagai T. 2011. Development of singleblastomere derived from two-cell embryos produced in vitroin pigs. Theriogenology, 76, 88-96Dai S J, Xu C L, Wang J, Sun Y P, Chian R C. 2012. Effect ofculture medium volume and embryo density on early mouseembryonic development: Tracking the development of theindividual embryo. Journal of Assisted Reproduction andGenetics, 29, 617-623Fraga M F, Ballestar E, Paz M F, Ropero S, Setien F, BallestarM L, Heine-Suñer D, Cigudosa J C, Urioste M, Benitez J,Boix-Chornet M, Sanchez-Aguilera A, Ling C, CarlssonE, Poulsen P, Vaag A, Stephan Z, Spector T D, Wu Y Z,Plass C, et al. 2005. Epigenetic differences arise during thelifetime of monozygotic twins. Proceedings of the NationalAcademy of Sciences of the United States of America, 102,10604-10609Fujita T, Umeki H, Shimura H, Kugumiya K, Shiga K. 2006.Effect of group culture and embryo-culture conditionedmedium on development of bovine embryos. Journal ofReproduction and Development, 52, 137-142Goovaerts I G, Leroy J L, Rizos D, Bermejo-Alvarez P,Gutierrez-Adan A, Jorssen E P, Bols P E. 2011. Single invitro bovine embryo production: Coculture with autologouscumulus cells, developmental competence, embryoquality and gene expression profiles. Theriogenology, 76,1293-1303Gopichandran N, Leese H J. 2006. The effect of paracrine/autocrine interactions on the in vitro culture of bovinepreimplantation embryos. Reproduction, 131, 269-277Haque F N, Gottesman I I, Wong A H. 2009. Not reallyidentical: Epigenetic differences in monozygotic twins andimplications for twin studies in psychiatry. American Journalof Medical Genetics (Part C: Seminars in Medical Genetics),151, 136-141Heyman Y, Vignon X, Chesné P, Le Bourhis D, Marchal J,Renard J P. 1998. Cloning in cattle: from embryo splittingto somatic nuclear transfer. Reproduction, Nutrition,Development, 38, 595-603Illmensee K, Levanduski M. 2010. Embryo splitting. Middle EastFertility Society Journal, 15, 57-63Joo B S, Kim M K, Na Y J, Moon H S, Lee K S, Kim H D.2001. The mechanism of action of coculture on embryodevelopment in the mouse model: direct embryo-to-cellcontact and the removal of deleterious components. Fertilityand Sterility, 75, 193-199Jung T, Fischer B. 1988. Correlation between diameter and DNAor protein synthetic activity in rabbit blastocysts. Biology ofReproduction, 39, 1111-1116Katayama M, Ellersieck M R, Roberts R M. 2010. Developmentof monozygotic twin mouse embryos from the time ofblastomere separation at the two-cell stage to blastocyst.Biology of Reproduction, 82, 1237-1247Kaminsky Z A, Tang T, Wang S C, Ptak C, Oh G H, Wong A H,Feldcamp L A, Virtanen C, Halfvarson J, Tysk C, McRae A F,Visscher P M, Montgomery G W, Gottesman I I, Martin N G,Petronis A. 2009. DNA methylation profiles in monozygoticand dizygotic twins. Nature Genetics, 41, 240-245Kelly S J. 1975. Studies on the potency of the early cleavageblastomere of the mouse. In: Balls M, Wild A E, eds., TheEarly Development of Mammals. Cambrige UniversityPress, Cambrige, London, New York, Melboure. pp. 97-105Linares T, King W A. 1980. Morphological study of the bovineblastocyst with phase contrast microscopy. Theriogenology,14, 123-133Loskutoff N, Johnson W, Betteridge K. 1993. The developmentalcompetence of bovine embryos with reduced cell numbers.Theriogenology, 39, 95-107Lewis N E, Rossant J. 1982. Mechanism of size regulation inmouse embryo aggregates. Journal of Embryology andExperimental Morphology, 72, 169-181Maluf M, Perin P M, Foltran Januário D A, Nascimento SaldivaP H. 2009. In vitro fertilization, embryo development, andcell lineage segregation after pre- and/or postnatal exposureof female mice to ambient fine particulate matter. Fertilityand Sterility, 92, 1725-1735Mitalipov S M, Yeoman R R, Kuo H C, Wolf D P. 2002.Monozygotic twinning in rhesus monkeys by manipulationof in vitro-derived embryos. Biology of Reproduction, 66,1449-1455Nagai M, Hori N, Miyamoto M, Sakaguchi M, Hayakawa Y,Kawai M, Kita M, Furuya T, Imai K. 2013. Effect of coculturewith intact embryos on development of bovine separateblastomere. Animal Science Journal, 84, 461-465Oppenheim S M, Moyer A L, Bondurant R H, Rowe J D,Anderson G B. 2000. Successful pregnancy in goats carryingtheir genetically identical conceptus. Theriogenology, 54,629-639Parikh F R, Nadkarni S G, Naik N J, Naik D J, UttamchandaniS A. 2006. Cumulus coculture and cumulus-aided embryotransfer increases pregnancy rates in patients undergoingin vitro fertilization. Fertility and Sterility, 86, 839-847Reichelt B, Niemann H. 1994. Generation of identical twinpiglets following bisection of embryos at the morula andblastocyst stage. Journal of Reproduction and Fertility,100, 163-172Rosenkrans Jr CF, First N L. 1994. Effect of free amino acidsand vitamins on cleavage and developmental rate of bovinezygotes in vitro. Journal of Animal Science, 72, 434-437Seike N, Saeki K, Utaka K, Sakai M, Takakura R, Nagao Y,Kanagawa H. 1989. Production of bovine identical twinsvia transfer of demiembryos without zonae pellucidae.Theriogenology, 32, 211-220Simón C, Mercader A, Garcia-Velasco J, Nikas G, Moreno C,Remohí J, Pellicer A. 1999. Coculture of human embryoswith autologous human endometrial epithelial cells inpatients with implantation failure. Journal of Clinical Endocrinology and Metabolism, 84, 2638-2646Spandorfer S D, Pascal P, Parks J, Clark R, Veeck L, Davis OK, Rosenwaks Z. 2004. Autologous endometrial coculturein patients with IVF failure: outcome of the first 1,030 cases.Journal of Reproductive Medicine, 49, 463-467Tagawa M, Matoba S, Narita M, Saito N, Nagai T, Imai K. 2008.Production of monozygotic twin calves using the blastomereseparation technique and Well of the Well culture system.Theriogenology, 69, 574-582Tao T, Niemann H. 2000. Cellular characterization of blastocystsderived from rabbit 4-, 8- and 16-cell embryos and isolatedblastomere cultured in vitro. Human Reproduction, 15,881-889Tao T, Reichelt B, Niemann H. 1995. Ratio of inner cell massand trophoblastic cells in demi- and intact pig embryos.Journal of Reproduction and Fertility, 104, 251-258Thouas G A, Korfiatis N A, French A J, Jones G M, Trounson AO. 2001. Simplified technique for differential staining of innercell mass and trophectoderm cells of mouse and bovineblastocysts. Reproductive Biomedicine Online, 3, 25-29Tsunoda Y, Tokunaga T, Okubo Y, Sugie T. 1987. Beneficialeffect of agar for the frozen storage of bisected embryos.Theriogenology, 28, 317-322Whitelaw N C, Whitelaw E. 2006. How lifetimes shapeepigenotype within and across generations. HumanMolecular Genetics, 15, 131-137Willadsen S M. 1981. The developmental capacity of blastomerefrom 4- and 8-cell sheep embryos. Journal of EmbryologyExperimental Morphology, 65, 165-172Willadsen S M, Godke R A. 1984. A simple procedure for theproduction of identical sheep twins. Veterinary Record,114, 240-243Williams T J, Elsden R P, Seidel Jr G E. 1984. Pregnancyrates with bisected bovine embryos. Theriogenology, 22,521–531. |
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