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• W1985702944 abstract "ObjectiveTo determine the cause of failed fertilization in mature eggs in IVF and ICSI cycles.DesignA prospective study of all IVF and ICSI patients from 1/1/2003 - 12/31/2004 who had less than a 40% fertilization rate per mature egg.Materials and methodsEggs were assessed for fertilization 17-20 hours post-insemination and re-assessed 23 hours post-insemination for any delayed fertilizations. If the percent of fertilized eggs was <40% of the total number of mature eggs, the unfertilized eggs were stained with 10 μg/ml Hoechst 33342 for 10 minutes at 37°C. The eggs were then washed and observed using phase contrast and fluorescence microscopy with an excitation wavelength of 355 nm and an emission wavelength of 465 nm. Observations included: the location and status (condensed or decondensed) of the sperm chromatin and presence or absence of maternal chromatin in the egg and polar body. When ≥50% of a patient’s (pt’s) eggs had a similar staining result, the result was considered conclusive and these pts were categorized based on the staining results. Pregnancies include pts who have on-going clinical pregnancies or who have delivered. Cycles in which pts used donor gametes were not included in the subsequent cycles.ResultsTabled 1ConclusionHoechst staining of unfertilized IVF eggs is helpful in identifying the putative cause of failed fertilization and enables physicians to provide some pts with a more definitive reason for lack of fertilization and future prognosis. Of the IVF pts with conclusive staining results, over 70% were the result of the sperm either not visible or in the PV space; most likely a defect in sperm binding to the zona pellucida or in fusing with the plasma membrane. These pts had a good prognosis with a 55.1% PR within 4 subsequent cycles. Of the ICSI pts with conclusive staining results, 32.4% were the result of the sperm either not visible or in the PV space. Technical problems such as a hyperelastic plasma membrane were the most likely cause for these unfertilized eggs. These pts had a good prognosis with a 50.0% PR within 2 subsequent cycles. Both IVF and ICSI pts whose staining results showed sperm in the ooplasm appear to have a poorer prognosis with only a 23.3% PR. Similarly, only 20% of pts whose eggs had a meiotic error have on-going pregnancies. ObjectiveTo determine the cause of failed fertilization in mature eggs in IVF and ICSI cycles. To determine the cause of failed fertilization in mature eggs in IVF and ICSI cycles. DesignA prospective study of all IVF and ICSI patients from 1/1/2003 - 12/31/2004 who had less than a 40% fertilization rate per mature egg. A prospective study of all IVF and ICSI patients from 1/1/2003 - 12/31/2004 who had less than a 40% fertilization rate per mature egg. Materials and methodsEggs were assessed for fertilization 17-20 hours post-insemination and re-assessed 23 hours post-insemination for any delayed fertilizations. If the percent of fertilized eggs was <40% of the total number of mature eggs, the unfertilized eggs were stained with 10 μg/ml Hoechst 33342 for 10 minutes at 37°C. The eggs were then washed and observed using phase contrast and fluorescence microscopy with an excitation wavelength of 355 nm and an emission wavelength of 465 nm. Observations included: the location and status (condensed or decondensed) of the sperm chromatin and presence or absence of maternal chromatin in the egg and polar body. When ≥50% of a patient’s (pt’s) eggs had a similar staining result, the result was considered conclusive and these pts were categorized based on the staining results. Pregnancies include pts who have on-going clinical pregnancies or who have delivered. Cycles in which pts used donor gametes were not included in the subsequent cycles. Eggs were assessed for fertilization 17-20 hours post-insemination and re-assessed 23 hours post-insemination for any delayed fertilizations. If the percent of fertilized eggs was <40% of the total number of mature eggs, the unfertilized eggs were stained with 10 μg/ml Hoechst 33342 for 10 minutes at 37°C. The eggs were then washed and observed using phase contrast and fluorescence microscopy with an excitation wavelength of 355 nm and an emission wavelength of 465 nm. Observations included: the location and status (condensed or decondensed) of the sperm chromatin and presence or absence of maternal chromatin in the egg and polar body. When ≥50% of a patient’s (pt’s) eggs had a similar staining result, the result was considered conclusive and these pts were categorized based on the staining results. Pregnancies include pts who have on-going clinical pregnancies or who have delivered. Cycles in which pts used donor gametes were not included in the subsequent cycles. ResultsTabled 1 ConclusionHoechst staining of unfertilized IVF eggs is helpful in identifying the putative cause of failed fertilization and enables physicians to provide some pts with a more definitive reason for lack of fertilization and future prognosis. Of the IVF pts with conclusive staining results, over 70% were the result of the sperm either not visible or in the PV space; most likely a defect in sperm binding to the zona pellucida or in fusing with the plasma membrane. These pts had a good prognosis with a 55.1% PR within 4 subsequent cycles. Of the ICSI pts with conclusive staining results, 32.4% were the result of the sperm either not visible or in the PV space. Technical problems such as a hyperelastic plasma membrane were the most likely cause for these unfertilized eggs. These pts had a good prognosis with a 50.0% PR within 2 subsequent cycles. Both IVF and ICSI pts whose staining results showed sperm in the ooplasm appear to have a poorer prognosis with only a 23.3% PR. Similarly, only 20% of pts whose eggs had a meiotic error have on-going pregnancies. Hoechst staining of unfertilized IVF eggs is helpful in identifying the putative cause of failed fertilization and enables physicians to provide some pts with a more definitive reason for lack of fertilization and future prognosis. Of the IVF pts with conclusive staining results, over 70% were the result of the sperm either not visible or in the PV space; most likely a defect in sperm binding to the zona pellucida or in fusing with the plasma membrane. These pts had a good prognosis with a 55.1% PR within 4 subsequent cycles. Of the ICSI pts with conclusive staining results, 32.4% were the result of the sperm either not visible or in the PV space. Technical problems such as a hyperelastic plasma membrane were the most likely cause for these unfertilized eggs. These pts had a good prognosis with a 50.0% PR within 2 subsequent cycles. Both IVF and ICSI pts whose staining results showed sperm in the ooplasm appear to have a poorer prognosis with only a 23.3% PR. Similarly, only 20% of pts whose eggs had a meiotic error have on-going pregnancies." @default.
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• W1985702944 title "Analysis of Failed Fertilization in IVF and ICSI Patients Using the DNA Specific Stain Hoechst 33342" @default.
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