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P-302 The development of one pronucleus “zygotes” is abnormal
P-301
P-302
O o c y t e s With O p a q u e C y t o p l a s m H a v e R e d u c e d Fertilization and Cleavage Rates Without a Significant I m p a c t o n C l i n i c a l P r e g n a n c y R a t e s . 1'2A. J. Carrillo,
T h e D e v e l o p m e n t o f O n e P r o n u c l e u s "Zygotes" Is A b n o r m a l . W.-S. A. Wun, C. Valdes, R. Dunn, C.-C. Wun,
1C. L. Cook, 2B. Lane, 1D. P r i d h a m , 2p. Risch, 2I. H. Silv e r m a n . 1Dept. of OB/GYN U n i v e r s i t y of Louisville a n d 2Fertility Center, A l l i a n t H e a l t h System, Louisville, KY.
Objectives: In some atypical occurrence, one pronucleus can only be observed in a n i n s e m i n a t e d oocyte. M u n e et al (1994 & 1995) r e p o r t e d cleaving one pronucleus "zygote" u s u a l l y h a d n o r m a l chromosome status. This s t u d y is to e x a m i n e d w h e t h e r one pronucleus "zygotes" develop norm a l l y or not. Design: This is a retrospective s t u d y in a clinic b a s e d A.R.T. program. All A.R.T. cases from J u n e , 1 9 9 6 - J a n u ary, 1997, were included. M a t e r i a l s and Methods: The fertilized oocytes t h a t did not get frozen or t r a n s f e r r e d were k e p t in culture till day 7 after egg retrieval. B l a s t u l a t i o n was t h e outcome examined a n d Chi S q u a r e t e s t was used for analysis. Two sample t-test was used for age comparison. Results:
Objectives: I n our p r o g r a m we frequently retrieve mat u r e a p p e a r i n g oocytes which have a n opaque region in the cytoplasm. This opaque region h a s a g r a n u l a r a p p e a r a n c e which m a y d i s a p p e a r after 2 - 3 days in culture. W h e n t h e s e oocytes fertilize, the pronuclei often a p p e a r w i t h i n this opaque region. This is in c o n t r a s t to t h e other m a t u r e oocytes which have a t r a n s p a r e n t cytoplasm. This s t u d y was designed to d e t e r m i n e t h e fertilization a n d cleavage r a t e of these opaque oocytes a n d t h e i r influence on clinical p r e g n a n c y rates. Design: Retrospective s t u d y of fertilization, cleavage a n d p r e g n a n c y r a t e s in a h o s p i t a l b a s e d IVF program. M a t e r i a l s a n d Methods: Oocytes from 225 p a t i e n t s retrieved in our F e r t i l i t y C e n t e r from Oct. 1994 to J a n . 1997 were studied. P a t i e n t m a n a g e m e n t a n d IVF lab procedures were t h e s a m e as described previously by us (52nd Ann. A S R M M e e t i n g 1996;Abs. #P-255). M a t u r e oocytes w i t h a n y opaqueness in t h e c y t o p l a s m were scored as having opaque cytoplasm (OC). E m b r y o s w i t h t h e g r e a t e s t n u m b e r of b l a s t o m e r e s a n d l e a s t a m o u n t of f r a g m e n t a t i o n r e g a r d l e s s of t h e oocyte of origin, were selected for r e t u r n 4 8 - 7 2 hours after retrieval. Clinical p r e g n a n c i e s was confirmed by u l t r a s o u n d . Results: Out of 225 p a t i e n t s , 158 (70%) h a d one or more oocytes w i t h a OC.
Patients
Fertilization Rate
Cleavage Rate
Clinical Pregnancy Rate
Control. OC
67% (379/565) 57% (1060/1865)*
94% (355/379) 89% (944/1060)**
33% (22/67) 35% (55/158)
* p < 0.001 vs control. ** p < 0.02 vs control; Chi Square test. The OC oocytes in the OC p a t i e n t group h a d a fertilization r a t e of only 32% (174) ~39 while t h e i r n o r m a l cohort oocytes in those s a m e p a t i e n t s h a d a fertilization r a t e of 67% ~1326]' ~8s6 There was no correlation b e t w e e n p e r c e n t of embryos developing from t h e OC oocytes and p r e g n a n c y r a t e in the OC patients. T h e r e were no significant differences bet w e e n t h e control p a t i e n t s a n d p a t i e n t s with OC oocytes in t h e following categories: age, diagnosis, i m p l a n t a t i o n r a t e a n d n u m b e r of F S H / h M G vials, oocytes a s p i r a t e d a n d embryos r e t u r n e d p e r patient. Conclusion: These d a t a d e m o n s t r a t e t h a t oocytes w i t h opaque c y t o p l a s m h a v e a m a r k e d l y reduced fertilization r a t e a n d slightly reduced cleavage rate. However, the presence of t h e s e oocytes does not reduce t h e chances for achieving a pregnancy. We do not know the cause or significance of t h e s e oocytes with opaque cytoplasm.
G. M. G r u n e r t . OB & GYN Associate, Houston, TX.
# Case
Age
Blast. %
Analysis
1. 2 pronuclei
66
2. i pronucleus
12
33.2 a (25-43) ~ 34.3 b (24-41)
46.5%c (322) t 0% d (21)
a vs. b Not significant ° vs. d P < 0.001
r: The range of age; t: Total number of pre-embryos Discussion: Most of the one pronucleus "zygotes" arr e s t e d d e v e l o p m e n t b e t w e e n 4 - 8 cell stage. It looked t h a t t h e y h a d problem to progress t h r o u g h the embryonic genome activation. "Zygotes" d i s p l a y i n g one pronucleus is not a n accident of norm. I t discloses defect(s) in pronucleus formation. The one pronucleus "zygote" is a b n o r m a l a n d no potential.
P-303 I S o l a t e T M As A N e w M e t h o d o f S p e r m S e p a r a t i o n . S . M . Tarchala, M. M. F a h y , H. M. H a u s e r m a n , K. Kallm a n n , K. Volentine, E. R a d w a n s k a , Z. Binor, M. W. Molo, R. G. Rawlins. Dept. Ob/Gyn, R u s h Medical Center, Chicago, IL. Objectives: The s e p a r a t i o n of h u m a n s p e r m a t o z o a from s e m i n a l p l a s m a is e s s e n t i a l prior to use in a s s i s t e d reproductive technologies. Effective m e t h o d s of s p e r m processing r e s u l t in a highly motile, morphologically n o r m a l s p e r m population containing m i n i m a l c o n t a m i n a t i o n from debris, b a c t e r i a a n d macrophages. The purpose of this s t u d y was to t e s t the efficacy of ISolate TM (a colloidal suspension of silica particles stabilized w i t h covalently bound hydrophilic silane) c o m p a r e d to a n o t h e r m e t h o d of s p e r m separation, Percoll TM (polyvinylpyrillidone coated beads, which h a s since been removed from t h e commercial m a r ket) with respect to yield, overall motility, forward progression, viability a n d % n o r m a l forms in two s p e r m populations, n o r m o s p e r m i c a n d compromised (thawed cryopreserved samples). Abstracts
$237
P-302
O o c y t e s With O p a q u e C y t o p l a s m H a v e R e d u c e d Fertilization and Cleavage Rates Without a Significant I m p a c t o n C l i n i c a l P r e g n a n c y R a t e s . 1'2A. J. Carrillo,
T h e D e v e l o p m e n t o f O n e P r o n u c l e u s "Zygotes" Is A b n o r m a l . W.-S. A. Wun, C. Valdes, R. Dunn, C.-C. Wun,
1C. L. Cook, 2B. Lane, 1D. P r i d h a m , 2p. Risch, 2I. H. Silv e r m a n . 1Dept. of OB/GYN U n i v e r s i t y of Louisville a n d 2Fertility Center, A l l i a n t H e a l t h System, Louisville, KY.
Objectives: In some atypical occurrence, one pronucleus can only be observed in a n i n s e m i n a t e d oocyte. M u n e et al (1994 & 1995) r e p o r t e d cleaving one pronucleus "zygote" u s u a l l y h a d n o r m a l chromosome status. This s t u d y is to e x a m i n e d w h e t h e r one pronucleus "zygotes" develop norm a l l y or not. Design: This is a retrospective s t u d y in a clinic b a s e d A.R.T. program. All A.R.T. cases from J u n e , 1 9 9 6 - J a n u ary, 1997, were included. M a t e r i a l s and Methods: The fertilized oocytes t h a t did not get frozen or t r a n s f e r r e d were k e p t in culture till day 7 after egg retrieval. B l a s t u l a t i o n was t h e outcome examined a n d Chi S q u a r e t e s t was used for analysis. Two sample t-test was used for age comparison. Results:
Objectives: I n our p r o g r a m we frequently retrieve mat u r e a p p e a r i n g oocytes which have a n opaque region in the cytoplasm. This opaque region h a s a g r a n u l a r a p p e a r a n c e which m a y d i s a p p e a r after 2 - 3 days in culture. W h e n t h e s e oocytes fertilize, the pronuclei often a p p e a r w i t h i n this opaque region. This is in c o n t r a s t to t h e other m a t u r e oocytes which have a t r a n s p a r e n t cytoplasm. This s t u d y was designed to d e t e r m i n e t h e fertilization a n d cleavage r a t e of these opaque oocytes a n d t h e i r influence on clinical p r e g n a n c y rates. Design: Retrospective s t u d y of fertilization, cleavage a n d p r e g n a n c y r a t e s in a h o s p i t a l b a s e d IVF program. M a t e r i a l s a n d Methods: Oocytes from 225 p a t i e n t s retrieved in our F e r t i l i t y C e n t e r from Oct. 1994 to J a n . 1997 were studied. P a t i e n t m a n a g e m e n t a n d IVF lab procedures were t h e s a m e as described previously by us (52nd Ann. A S R M M e e t i n g 1996;Abs. #P-255). M a t u r e oocytes w i t h a n y opaqueness in t h e c y t o p l a s m were scored as having opaque cytoplasm (OC). E m b r y o s w i t h t h e g r e a t e s t n u m b e r of b l a s t o m e r e s a n d l e a s t a m o u n t of f r a g m e n t a t i o n r e g a r d l e s s of t h e oocyte of origin, were selected for r e t u r n 4 8 - 7 2 hours after retrieval. Clinical p r e g n a n c i e s was confirmed by u l t r a s o u n d . Results: Out of 225 p a t i e n t s , 158 (70%) h a d one or more oocytes w i t h a OC.
Patients
Fertilization Rate
Cleavage Rate
Clinical Pregnancy Rate
Control. OC
67% (379/565) 57% (1060/1865)*
94% (355/379) 89% (944/1060)**
33% (22/67) 35% (55/158)
* p < 0.001 vs control. ** p < 0.02 vs control; Chi Square test. The OC oocytes in the OC p a t i e n t group h a d a fertilization r a t e of only 32% (174) ~39 while t h e i r n o r m a l cohort oocytes in those s a m e p a t i e n t s h a d a fertilization r a t e of 67% ~1326]' ~8s6 There was no correlation b e t w e e n p e r c e n t of embryos developing from t h e OC oocytes and p r e g n a n c y r a t e in the OC patients. T h e r e were no significant differences bet w e e n t h e control p a t i e n t s a n d p a t i e n t s with OC oocytes in t h e following categories: age, diagnosis, i m p l a n t a t i o n r a t e a n d n u m b e r of F S H / h M G vials, oocytes a s p i r a t e d a n d embryos r e t u r n e d p e r patient. Conclusion: These d a t a d e m o n s t r a t e t h a t oocytes w i t h opaque c y t o p l a s m h a v e a m a r k e d l y reduced fertilization r a t e a n d slightly reduced cleavage rate. However, the presence of t h e s e oocytes does not reduce t h e chances for achieving a pregnancy. We do not know the cause or significance of t h e s e oocytes with opaque cytoplasm.
G. M. G r u n e r t . OB & GYN Associate, Houston, TX.
# Case
Age
Blast. %
Analysis
1. 2 pronuclei
66
2. i pronucleus
12
33.2 a (25-43) ~ 34.3 b (24-41)
46.5%c (322) t 0% d (21)
a vs. b Not significant ° vs. d P < 0.001
r: The range of age; t: Total number of pre-embryos Discussion: Most of the one pronucleus "zygotes" arr e s t e d d e v e l o p m e n t b e t w e e n 4 - 8 cell stage. It looked t h a t t h e y h a d problem to progress t h r o u g h the embryonic genome activation. "Zygotes" d i s p l a y i n g one pronucleus is not a n accident of norm. I t discloses defect(s) in pronucleus formation. The one pronucleus "zygote" is a b n o r m a l a n d no potential.
P-303 I S o l a t e T M As A N e w M e t h o d o f S p e r m S e p a r a t i o n . S . M . Tarchala, M. M. F a h y , H. M. H a u s e r m a n , K. Kallm a n n , K. Volentine, E. R a d w a n s k a , Z. Binor, M. W. Molo, R. G. Rawlins. Dept. Ob/Gyn, R u s h Medical Center, Chicago, IL. Objectives: The s e p a r a t i o n of h u m a n s p e r m a t o z o a from s e m i n a l p l a s m a is e s s e n t i a l prior to use in a s s i s t e d reproductive technologies. Effective m e t h o d s of s p e r m processing r e s u l t in a highly motile, morphologically n o r m a l s p e r m population containing m i n i m a l c o n t a m i n a t i o n from debris, b a c t e r i a a n d macrophages. The purpose of this s t u d y was to t e s t the efficacy of ISolate TM (a colloidal suspension of silica particles stabilized w i t h covalently bound hydrophilic silane) c o m p a r e d to a n o t h e r m e t h o d of s p e r m separation, Percoll TM (polyvinylpyrillidone coated beads, which h a s since been removed from t h e commercial m a r ket) with respect to yield, overall motility, forward progression, viability a n d % n o r m a l forms in two s p e r m populations, n o r m o s p e r m i c a n d compromised (thawed cryopreserved samples). Abstracts
$237