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Blastocyst stage microarray-based genomic screening in improving clinical pregnancy rate and implantation rate

Blastocyst stage microarray-based genomic screening in improving clinical pregnancy rate and implantation rate
AuthorHsing-Hua Lai, Huai-Ling Wang, Wen-Yi Chiang, Shih-Chieh Huang, Ya-Hui Hsu
EditorShih-Chieh Huang
Seminar name2011 TSRM
KeywordPGS, aCGH, blastocyst culture
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Identification of embryos with the highest implantation potential is a crucial step in IVF. Blastocyst transfer (BT) has been proved to have higher implantation rate and live birth rate than D3 embryo transfer owing to a further embryo selection mechanism. However, morphological grading provides us only part of clue to choosing a best quality blastocyst. As chromosome abnormality plays a role in pregnancy failure for woman of advanced maternal age and repeated implantation failure, it is important to evaluate the chromosome status before BT to reduce miscarriage rate.


Materials and Methods

Patient Profile
Patient no.34
Average age36.9
Average age (+OD)31.1
Average previous IVF2.76
Blastocysts biopsied228
Warmed BT cycles23
Trophectoderm Biopsy
》On day 4 of embryonic development, the zona pellucida was breached using a laser and transferred to blastocyst medium.
》On day 5 or 6, expanding blastocysts underwent biopsy of herniating trophectoderm cells.
》Aspirated cells were froze at -20℃ and subjected to chromosome analysis with array comparative genomic hybridization (aCGH).
》Biopsied blastocysts were vitrified using Cryotop method.
》Biopsy procedure

Array comparative genomic hybridization (aCGH)


Results

Chromosome Analysis
》Two hundred and twenty-eight blastocysts were analyzed. The results were as follows:
no.Ratio
Euploidy11952%
Monosomy3114%
Trisomy2812%
Multiple aneuploidy2611%
No signal2411%

》Although prolonged culture has been accomplished, there were still about 50% blastocysts possessed chromosomal abnormality
Blastocyst Morphology and Aneuploidy Rate
》To examine the relationship of chromosome status and the morphology of blastocyst, aneuploidy rate were analyzed according to the grading of the inner cell mass and trophectoderm, respectively.
ICMno.Aneuploidy rate
A2642%
B17240%
C00%
TENo.Aneuploidy rate
A1547%
B11641%
C6737%

》Aneuploidy rate were analyzed according to the blastocyst grading.
Gradingno.Aneuploidy rate
AA450%
AB1947%
AC30%
BA1145%
BB9745%
BC6439%
Blastocyst683%

》High aneuploidy rate was observed in blastocyst with lower growth rate.
》No obvious relationship between blastocyst morphology and aneuploidy rate.
Age and Aneuploidy Rate
》Patients were classified as 4 groups according to their age to examine the relationship of age and aneuploidy rate.
Age#≦3031~3536~39≧40
Case No.15883
Emb. No.86524819
Aneuploidy Rate23%56%58%63%
#: adopting donor’s age for analysis

》As previous report, high aneuploidy rate is related advanced maternal age. Significant differences were observed among ≦30 group and every other group.
Clinical Outcome
》Warmed BT cycles with aCGH (aCGH group) were compared with contemporary group, which aCGH was not performed, and the clinical outcome was listed below:
aCGH groupContemporary group (non-aCGH)
Vitrification-warmed BT cycles23111
Average blastocysts transferred/BT1.5***(34/23)2.3(251/111)
Clinical pregnancy rate(cycles producing gestational sac/BT)74% (17/23)61%(68/111)
Implantation rate(fetal heartbeats/transferred embryo)50%*(17/34)31%(77/251)
Multiple pregnancy rate(multiple pregnancy /pregnancy cycle)12%(2/17)24%(16/66)
》In aCGH group, fewer blastocysts were transferred per cycle, resulted in a relatively low multiple pregnancy rate.
》Clinical pregnancy rate was slightly higher in aCGH group, though without significant difference.
Clinical Outcome in 3 aCGH Sub-group
》In aCGH group, patients were grouped into 3 sub-sets: single embryo transfer (eSET), double embryo transfer (DET) and triple embryo transfer (BT).
eSETDETBT
Cycles1391
Clinical pregnancy rate(cycles producing gestational sac/BT)62%(8/13)89%(8/9)100%(1/1)
Implantation rate(fetal heartbeats/transferred embryo)54%(7/13)44%(8/18)67%(2/3)
Multiple pregnancy rate(multiple pregnancy /pregnancy cycle)0%(0/8)13%(1/8)100%(1/1)
》In eSET cycles, pregnancy rate could be achieved by means of aCGH in combination with BT.
》Multiple pregnancy rate was successfully reduced in eSET group.
Conclusions
Our preliminary data indicated that blastocyst stage preimplantation chromosome screening could improve implantation rate. Although there was no statistical significant, pregnancy rate was higher in aCGH group. More clinical data was needed for further study.
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