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Human embryo blastocyst culture and relative factors research

Author: LinXiaoNa
Tutor: ChenZuoBo
School: Zhejiang University
Course: Obstetrics and Gynaecology
Keywords: Blastula Cultured in vitro Blastocyst formation rate
CLC: R321
Type: Master's thesis
Year: 2001
Downloads: 110
Quote: 1
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Abstract

Background material: conventional in vitro fertilization - embryo transfer (In-vitro fertilization and embryotransfer, referred to IVF-ET) embryos cultured in vitro for 2 to 3 days, and transplanted back into the uterine cavity 4 to 8-cell stage embryo implantation rate of about 10 to 30 %. However, the embryos were cultured in vitro to the blastocyst stage transplantation more in line with physiological needs, and can reduce the number of embryos transferred, is conducive to select viable embryos beyond 4 to 8-cell stage development block, and can help repair micromanipulation, improve the implantation rate, to avoid one of the methods of fetal reduction. Blastocyst stage embryo culture co-culture (co-culture) and the sequential culture (sequential culture) two technologies. The former need to be prepared helper cells, complicated steps, and the risk of cross infection, sequential culture is relatively simple and feasible. Purpose: This topic shifted into surplus embryos in the first 3 days after transplantation default culture fluid (IVC-Three Blastocyst Medium, BlastocystMedium or HTF) for follow-up training, observe the blastocyst formation, analysis of the 3-day embryo cell number and quality the relationship between the classification and blastocyst formation, compare the three blastocyst formation rate of the culture medium, 5 days embryo transfer provide a theoretical basis for clinical. Materials and Methods: This topic select from November 1999 to April 2001 in our hospital volunteer intracytoplasmic receiving IVF-ET or intracytoplasmic sperm injection - embryo transfer (intracytoplasmic sperm injection and embryo transfer, referred to as ICSI-ET) 18 infertile patients received surplus embryos after transplantation 54. IVF-ET clinical routine, before transplantation at New York University, USA 1 * F center standard line embryo quality grading, record the number and level of embryonic cells. The remaining embryos to the default culture medium containing 10% synthetic serum substitute (SSS), the daily observation of embryonic growth and development until the degradation or seven days after fertilization, the 6th day of non-degraded replaced with fresh culture medium. Results: the 18 patients with an average age of 29.78 ± 3.34 years old, average egg retrieval 10,33 ± 5.78, 7.56 ± 3.76 average fertilization, an average of embryos transferred 3.39 ± 0. 50. 16 of 54 embryos formed blastocysts, total blastocyst formation rate of 29% stone; 12 blastocysts full expansion, thinning of the zona pellucida; 3 blastocyst hatching phenomenon. Six cases among the 18 cases of clinical pregnancy, clinical pregnancy rate n. ac, o, two cases of single births, z cases of twin, z cases of three children; total of 61 embryos transferred, planted 12 implantation rate of 19.7%. Discussion: Analysis of the first three days of the relationship between the embryo and blastocyst formation, blastocyst formation rate was found with the increase in the number of the first three days of embryonic cells increased, both were significantly positively correlated (0,99, P <0 knife 01 ), the number of cells in more than eight blastocyst formation rate of up to 50%. At the same time, the embryo quality level l to 2.5 level blastomere than 3 to 4 has a higher rate of blastocyst formation doors 4.90 9. lo examination was no significant difference (X B L.69, P> 0 knife 5). In a choice of three follow-up culture medium, IVC-Three Medium has the highest blastocyst formation rate compared with the other two culture medium were significantly different (expansion book GZ, P kiss knife 25), and standardized by the number of embryonic cells the after the IVC-Thlee of Medium blastocyst formation rate of 56.1%, similar to foreign reports figures. This paper also analyzes the remaining embryo blastocyst formation or absence of clinical pregnancy, 10 cases formed blastocysts were 4 cases of pregnancy, eight cases of blastocyst formation of two cases of pregnancy, no significant difference in the test, but the combination of reported that certain clinical significance. The domestic conventional IVF embryo transplantation 3 to 4, or even five, multiple pregnancy brings a lot of complications. Transplantation l-2 blastocysts avoid rabbit effective method of multiple births. Due to the small number of observed embryos, the blastocyst quality evaluation is still a lack of experience, needs further laboratory research. Blastocyst stage embryo culture and transfer there are some practical difficulties, such as culture transplantation cancellation rate increased blastocyst quality assessment standards confusion. Conclusion: 1. The rest of the embryo transplant follow-up training, can reach a certain number of blastocysts to prove that under certain laboratory conditions, the application of sequential culture blastocyst stage embryo culture technology surgery is feasible. 2, 3 days after fertilization, embryonic cells and the blastocyst formation rate was significantly positively correlated; different embryo quality classification of blastocyst formation rate was no significant difference, but specific figures vary greatly, there is a certain significance. Note 3 days in good shape 8-cell embryos can be cultured for more than three to five days, so that a more accurate selection of viable embryos. 3. The IVC-ThxeeMedium than the blastocyst Medium and HTF culture medium has a higher blastocyst formation rate, available for clinical use. 4. Transplant two blastocysts to avoid multiple pregnancy, an effective way to reduce complications.

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