MINISTRY OF SCIENCE AND TECHNOLOGY
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E-mail: ured@znanost.hr
Title: Steroidogenesis
Title: Placental hormonal function. Endocrinological events
during pregnancy.
Title: Serum estradiol and progesterone ration during induction
of ovulation relative to the success of in vitro fertilization
Title: Preincubation of sperm in milk impruves in vitro
fertilization outcome
Faculty: Medicinski fakultet Sveučilište u Zagrebu
Author: BARIŠIĆ DUBRAVKO
Date of defense: 12/19/94
Language: hrvatski
Number of pages: 84
Summary: To evaluate the efficacy of milk as medium for human sperm
processing, different experiments were performed and the results compared
with those obtained for TEST-yolk medium. Washed spermatozoa were divided
into two portions, and an equal volume of homogenized milk,heated to 95 C
for 10 min, was added to one portion, and TEST-yolk was added to the other.
After incubation at 5 C sperm were washed and evaluated for the ability to
undergo capacitation and acrosome reaction, for the effects on sperm
motility and movement characteristics, physical and functional integrity of
sperm memebrane, as well as for sperm fertilizing potential, as measured by
Sperm Penetration Assay (SPA), Hemizona Assay (HZA), and in vitro
fertilization (IVF). No significant (p<0.05) differences in percentage of
acrosome reacted sperm were noted following various incubation periods in
both media. Two-hour preincubation was therefore arbitrarily chosen as a
standard for further experiments. Sperm preincubated at 5 C in milk yielded
a significantly (p<0.05) higher percentage of acrosome reaction (25.1+-4.1)
than the sperm preincubated at 37 C in milk (13.6+-2.0). Statistically, no
difference in sperm motility and HOS positive sperm were observed when the
sperm were incubated either in regular milk (pH: 6.6) or pH adjusted milk
(pH: 7.4). Sperm motility and movement characteristics, except the
amplitude of lateral head displacement were significantly (p<0.05) lower
following preincubation in milk than in TEST-yolk. However, sperm
preincubated in milk yielded a significantly (p<0.001) higher penetration
rate in SPA (54.0+-8.6) than in TEST-yolk medium (13.9+-4.5). Penetration
indexes were not statistically different (1.4+-0.2 vs 1.1+-0.1). A higher
number of sperm was bound to zona pellucida, when the sperm were
preincubated in milk (101.8+-14.5)than in TEST-yolk (81.3+-7.8). In the
clinical study, 41 consecutive couples, undergoing an IVF program, were
randomized. Oocytes from one group of couples were inseminated with
milk-treated spermatozoa, and those from another group of couples were
inseminated with TEST-yolk-treated sperm. Seventy five out of 100 mature
oocytes (75%) were fertilized in group with milk treated sperm, and 45 out
of 64 (70%) in TEST-yolk group. The difference was not statistically
significant. Based on the results, and since milk is natural,
simple-to-prepare and readily available, it should be the medium of choice
for processing of human semen in Assisted Reproductive Technology
procedures.
Keywords: Sperm preincubation, Milk, TEST-yolk, In vitro fertilization
Title: Human sperm preincubation in milk improves the in vitro
fertilization
Faculty: Medicinski fakultet Sveučilište u Zagrebu
Mentor: SUCHANEK ERNEST
Date of defense: 12/19/94
Number of pages: 84
Author: Barišić Dr med., dr sc. Dubravko
Degree level: Ph.D.
Title: Correlation between follicular fluid hyaluronic acid and
in vitro fertilization of human oocytes
Institution: University of Rochester Medical Center
Year: 1992
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