سه شنبه 11 دی 1403
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دوره 5، شماره 2 - ( 4-1386 )
جلد 5 شماره 2 صفحات 22-17 |
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Khalili M A, Anvari M. The effect of in vitro culture on cleavage rates and morphology of the in vivo- developed embryos in mice. IJRM 2007; 5 (2) :17-22
URL: http://ijrm.ir/article-1-63-fa.html
URL: http://ijrm.ir/article-1-63-fa.html
The effect of in vitro culture on cleavage rates and morphology of the in vivo- developed embryos in mice. International Journal of Reproductive BioMedicine. 1386; 5 (2) :17-22
چکیده: (2934 مشاهده)
Background: Research studies on reproductive mechanism of laboratory animals are essential for further advancement of assisted reproductive techniques (ART). One of these studies includes the assessment of in-vitro development of pre-implantation embryos. The objective was to compare the cleavage rates and morphology of in-vivo formed 2 to 8 cell embryos and blastocysts with in-vitro culture of the same embryos for 24 h.
Materials and Methods: 6-8 weeks old female NMRI mice were superovulated with 8IU pregnant mare's serum gonadotropin (PMSG, ip). Two superovulated animals were caged with one male mouse for mating. Mated mice were killed by cervical dislocation at different time intervals to collect a total of 200 (50/ each) 2, 4, 8, and blastocyst embryos from uterine tubes and horns. Following morphological evaluation and cleavage rates, all embryos were incubated in Whittingham's T6 media+5% BSA for 24 h. Following incubation at 37ºC in 5% CO2, the cleavage rates as well as morphological feature of each embryo was re-evaluated and compared with the original embryos.
Results: The best quality embryos collected from uterine tubes were at 2-cells stage, which were reduced when compared with in-vivo developed 4-8 cells embryos. 88% and 52% of 2 and 8 cells embryos were respectively at grade A stage. 28 embryos out of 50 eight-cell embryos were at grades C and D after incubation. Following in vitro culture, the development of 16%, 24%, 24%, and 40% of the 2, 4, 8 cells, and blastocysts were arrested, respectively. Also, only 2 blastocysts (8%) reached the hatching stage which in comparison with in-vivo blstocysts were increased (P>0.05).
Conclusion: In-vitro culture of the in-vivo formed embryos reduced their cleavage rates and morphology, especially at more advanced stages. Therefore, it becomes necessary to improve the in-vitro culture condition and to transfer the embryos at early stage to consequently improve the implantation rates.
Materials and Methods: 6-8 weeks old female NMRI mice were superovulated with 8IU pregnant mare's serum gonadotropin (PMSG, ip). Two superovulated animals were caged with one male mouse for mating. Mated mice were killed by cervical dislocation at different time intervals to collect a total of 200 (50/ each) 2, 4, 8, and blastocyst embryos from uterine tubes and horns. Following morphological evaluation and cleavage rates, all embryos were incubated in Whittingham's T6 media+5% BSA for 24 h. Following incubation at 37ºC in 5% CO2, the cleavage rates as well as morphological feature of each embryo was re-evaluated and compared with the original embryos.
Results: The best quality embryos collected from uterine tubes were at 2-cells stage, which were reduced when compared with in-vivo developed 4-8 cells embryos. 88% and 52% of 2 and 8 cells embryos were respectively at grade A stage. 28 embryos out of 50 eight-cell embryos were at grades C and D after incubation. Following in vitro culture, the development of 16%, 24%, 24%, and 40% of the 2, 4, 8 cells, and blastocysts were arrested, respectively. Also, only 2 blastocysts (8%) reached the hatching stage which in comparison with in-vivo blstocysts were increased (P>0.05).
Conclusion: In-vitro culture of the in-vivo formed embryos reduced their cleavage rates and morphology, especially at more advanced stages. Therefore, it becomes necessary to improve the in-vitro culture condition and to transfer the embryos at early stage to consequently improve the implantation rates.
نوع مطالعه: Original Article |
فهرست منابع
1. Edwards RG. An introduction to Bourn Hall: The biomedical background of Bourn Hall Clinic. In: Textbook of in vitro fertilization and assisted reproduction; ed. Brinsden PR. Taylor & Francis Group; London 2005; 26: 1-8. [DOI:10.1201/b14680-2]
2. Khalili MA, Moinia F. Role of embryo morphology and cumulative embryo score in pregnancy outcome from in-vitro fertilization and intracytoplasmic sperm injection cycles. Mid East Fert Soc J 2002; 7: 231-236.
3. Barsiter BD. Mouse embryo cleavage, metabolism and viability: Role of medium composition. Hum Reprod 1993; 8: 288-295. [DOI:10.1093/oxfordjournals.humrep.a138039]
4. Ebner T, Moser M, Sommeryruber M, Gaiswinkler U. Presence, but not type of degree of extension of a cytoplasmic halo has a significant influence on preimplantation development and implantation behavior. Hum Reprod 2003; 18: 2406-2412. [DOI:10.1093/humrep/deg452]
5. Schleve LA, Wilcox LS. Use of assisted reproductive technology in United States, 1996 and 1998. JAMA 2002; 287: 1521-1522. [DOI:10.1001/jama.287.12.1521]
6. Olson SE, Seidel GE. Culture of in-vitro produced bovine embryos with vitamin E improves development in vitro and after transfer to recipients. Bio Reprod 2000; 62: 248-252. [DOI:10.1095/biolreprod62.2.248]
7. Kikuchi K, Kashiwazaki N, Noguchi J, Shimada A, Takahashi R, Hirabayashi M. Developmental competence, after transfer to recipients, of porcine oocyte matured, fertilized and cultured in vitro. Bio Reprod 1999; 60: 336-340. [DOI:10.1095/biolreprod60.2.336]
8. Aflalo ED, Sod-Moriah UA, Potashnik G, Har-Vardi I. Differences in the implantation rates of rat embryos developed in-vivo and in-vitro: possible role for plasminogen activators. Fert Steril 2004; 81: 780-785. [DOI:10.1016/j.fertnstert.2003.10.014]
9. Hogan B, Beddington R, Costantini F, Lacey E. Manipulating the mouse embryo: a laboratory manual. 2nd ed NY: Cold spring harbor laboratory, 1995.
10. Wolf DP. In vitro fertilization and embryo transfer. A manual of basic techniques. Penum Press; Phil, USA, 1988.
11. Rubio C, Simon C, Mercader A, Garcia-Velasco J, Remohi J, Pellicer A. Clinical experience employing co-culture of embryos with autologous human endometrial epithelial cells. Hum Reprod 2000; 15: 31-38.
12. Vander-Auwera I. Mouse embryo development in vivo. Embryo Mail News, 2000.
13. Braude P, Bolton V, Moore S. Human gene expression first occurs between the four and eight cell stage of pre-implantation development. Nature 1988; 332: 459-461. [DOI:10.1038/332459a0]
14. Gardner DK, Lane M. Amino acids and ammonium regulate mouse embryo development in culture. Bio Rep 1993; 48: 377-385. [DOI:10.1095/biolreprod48.2.377]
15. Carroll PM, Richards WG, Darrow AL, Wells JM, Strickland S. Pre-implantation mouse embryos express a cell surface receptor for tissue-plasminogen activator. Develop 1993; 119: 191-198.
16. Goto Y, Noda Y, Mori T, Nakano M. Increased generation of reactive oxygen species in embryo cultured in vitro. Free Rad Biol Med 1993; 15: 69-75. [DOI:10.1016/0891-5849(93)90126-F]
17. Feugang JM, Langendonckt AV, Sayoud H, Rees JF, Pampfer S, Moens A, et al. Effect of prooxidant agents added at the morula/blastocyst stage on bovine embryo development, cell death, and glutathione content. Zygote 2003; 11: 107-118. [DOI:10.1017/S0967199403002144]
18. Munne G. Embryo morphology, developmental rate and maternal age are correlated with chromosome abnormalities. Fert Steril 1995; 64: 382-390.
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