multipurpose handling media for iui? · multipurpose handling medium (mhm) for iui author: jayant...
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Multipurpose Handling Media for IUI?
Jayant G Mehta Director Sub-Fertility laboratory and Quality Control Manager
Queen’s Hospital
Romford
UK
IUI – Success Rates
Sperm Function dependent on the pH
Laboratory Equipment for IUI
pH – buffering range
Multipurpose Handling Medium
and H
Contains: MOPS and HEPES
Sperm Preparation for IUI
Preparation for intrauterine insemination (IUI)
is to obtain
– large number of motile sperm
– the most fertilizable population of sperm
DENSITY GRADIENT
New line of density gradient products have
been manufactured, specifically,
– colloidal silica particle-based density gradients
ISolate®
PureSperm®,
ISOLATE
Objectives of the Preliminary Study
Compare two sperm wash media:
– Quinn’s (Origio) and MHM (Irvine Scientific)
Compare 24 hrs survival
Pregnancy with MHM
Compare two gradient separation materials:
– ISolate (Irvine Scientific) and Puresperm (Nidacom)
Higher motile sperm yield
Design of the Study
Four arms to the study.
– 1. ISOLATE and MHM medium
– 2. ISOLATE and Quinn’s sperm wash medium
– 3. PURESPERM and MHM medium
– 4. PURESPERM and Quinn’s sperm wash
Recovered count
% motile count
% 24 hrs survival of progressive motile sperm
Inclusion Criteria
Semen sample > than 3 mls
Sperm count > 40 million per mL
Sperm progressive motility > 50%
Method
Sample allowed to be liquefied for 20 minutes
at 37 C
Discontinuous gradients prepared (40% and
80% of ISOLATE® and PURESPERM®)
Allowed to stand in a heating block at 37 C.
Sample divided into two equal parts
Each part minimum volume 1.5 mls
Mean Sperm Count Recovery (Mil/mL)
Age No of
Patients
(n=50)
Mean Count
Recovered
with
ISOLATE ±
SD
Mean Count
recovered
with
PURE
SPERM ± SD
% Pregnancy
After
insemination
using
ISOLATE and
MHM
25-30 13 12.5 ± 2.3 9.0 ± 2.6 15.4 (2/13)
30-35 20 17.8 ± 3.4 12.7± 3.2 25.0 (5/20)
36-40 12 17.3 ± 2.9 11.8 ± 2.1 8.3 (1/12)
>40 05 18.0 ± 3.7 11.0 ± 2.4 0.0 (0/5)
Mean % Motile Sperm Recovery – ISOLATE/24hrs survival
Age No of
Patients
(n=50)
Mean %
Motile
recovery
washed with
MHM (1)
± SD
Mean %
Motile
recovery
washed
with
Quinn’s (2)
± SD
Mean % 24
hrs
progressive
survival(1)
± SD
Mean % 24
hrs
progressive
survival(2)
± SD
25-30 13 90.8 ± 5.8 91.3 ± 7.2 74.5 ± 4.3 63.4 ± 8.2
30-35 20 88.4 ± 4.4 82.4 ± 5.6 70.3 ± 6.2 59.5 ± 7.4
36-40 12 90.1 ± 3.2 81.3 ± 6.6 77.4 ± 7.4 57.6 ± 6.5
>40 05 91.2 ± 3.6 84.2 ± 4.2 77.5 ± 6.4 58.7 ± 8.3
Mean % Motile Sperm Recovery PURESPERM/24hrs survival
Age No of
Patients
(N=50)
Mean %
Motile
recovery
washed
with MHM
(3) ± SD
Mean %
Motile
recovery
washed
with
Quinn’s
(4) ± SD
Mean 24
hrs
progressi
ve
survival
(3) ± SD
Mean 24
hrs
Progressi
ve
survival
(4) ± SD
25-30 13 89.6 ± 6.7 86.7 ± 4.7 72.7 ± 3.7 57.9 ± 5.9
30-35 20 88.8 ± 4.6 85.9 ± 5.1 70.3 ± 5.3 58.4 ± 6.2
36-40 12 88.2 ± 7.2 82.3 ± 6.9 72.5 ± 7.2 60.3 ± 7.2
>40 05 90.1 ± 4.6 80.2 ± 5.3 75.8 ± 4.6 58.6 ± 6.3
CONCLUSION
Statistical analysis not possible due to low
number of patients
Trend suggests:
– ISOLATE separation yields higher progressive
motile sperm
– MHM washed sperm had higher mean% of
progressive 24 hrs survival
– This combination resulted in pregnancies.
– Possible reason pH regulation with MOPS and
HEPES?
FUTURE
A larger study
Look at the influence of MHM on sperm
kinetics