Download - UM Research Repository

Impact of aspartate and serine on
mouse embryo development in vitro
Jaffar
1*
Ali
and Nafisa
2
Ali
1*Department
of Obstet. & Gynaecol, Faculty of Medicine, University of Malaya, Kuala
Lumpur, Malaysia; Email: [email protected]; Tel: +603 7949 6898; Fax: +603 7949 4193
Abstract No.
Aspir-0176
2School
of Archaeology & Anthropology, ANU College of Arts and Social Sciences, AD Hope
Building, The Australian National University, Canberra, ACT 2601, Australia
BACKGROUND
Attempts to improve in vitro culture conditions of early embryos
must determine the nutritional requirement of individual nutrient
components that impact embryo development in vitro. The
findings of the present research effort is to assist elucidate the
effect of two amino acids on embryo development. Amino acids
are crucial for protein synthesis, energy metabolism and for its
role as osmolytes.
Image 1: Human embryo
generated in synthetic proteinfree ECM; now a youngster aged
about 17 yrs.
CONCLUSION
Supplementation of both aspartate and serine in embryo culture medium
appear detrimental but these effects were insignificant. These responses
were obtained under the confounding influences of bovine serum albumin
that is present in conventional embryo culture media (ECM). The present
findings showed the addition of these two amino acids in conventional ECM
containing serum proteins may not be useful and may even be harmful. This
suggests the practice of supplementing ECM with aspartate and serine be
re-examined.
Table 1: Effect of Aspartate on embryo development in vitro
Concentration of
Aspartate (mM)
0
250
500
750
1000
Day 1 Embryos
79
76
79
77
76
A 15-20 YEARS RESEARCH EFFORT: This study is part of a
15-20 years of research effort to culture embryos in synthetic
and chemically defined conditions (Ali, 1997; Ali et al., 2000; Ali,
2007) devoid of added donor serum proteins.
Day 2
71
73
75
75
73
Day-3
40
37
48
42
37
Day-4
40
36
48
42
36
OBJECTIVE
Day-5
27
26
34
33
27
The objective is to determine the optimal concentrations,
tolerance levels and effects of aspartate and serine on mouse
embryo development in vitro.
Day-6
%Dead r=0.4093
(p=1)
p-value Chi-square for
trend+
23
26
33
31
25
29.1
34.2
41.8
40.3
32.9
0.9901
0.8672
0.9318
0.9944
MATERIALS AND METHODS
Earle’s Basal salt solution was supplemented with 5mg/ml of
bovine serum albumin, antibiotics penicillin/ streptomycin sulfate,
and pyruvate/ lactate. Treatments consisted of culture media
supplemented with graded amounts (0, 250,500,750 and
1000mM) of aspartate and serine. Quakenbush sibling mice
zygotes were randomly apportioned to individual treatments. The
numbers of replicates were 5 and 4 respectively. The end point
was the development of day 6 (expanded, hatching, hatched)
embryos.
Replicates=5; n= 387 embryos
+Treatments compared individually with control
Table 2: Effect of Serine on embryo development in vitro
Concentration of
Serine (mM)
Day 1 Embryos
RESULTS
The proportion of day 6 embryos developed decreased when
supplemented with aspartate with the portion of dead increased
29.1, 34.2, 41.8, 40.3, 32.9% with the addition of graded
amounts (0, 250,500,750 and 1000mM respectively) of aspartate
with maximum effect seen with treatment supplemented with
500mM but this was not significant. Likewise all treatments
supplemented with serine resulted in insignificant increases in
embryonic death and was harmful at all concentrations tested
(%dead: c =51.6, 250mM=58.3%, 500mM=53.3%, 750mM=75%
and 1000mM=61.7% respectively; r = 0.6271; p=0.2575).
REFERENCE
Ali,1997. FSA Annual Conference, Adelaide, Australia
Ali et al., 2000, Human Reprod 15:145-156
Ali, 2004, MEFS J. 9(2):118-127
Ali, J. United States Patent US2009/0226879 A1;Dec 2007;
PCT 2008 (No. US2009/0226879 A1); (PCT US No. PCT/US2008087818)
0
250
500
750
1000
62
60
60
60
60
57
56
57
55
55
40
36
40
31
31
40
33
33
28
31
39
32
28
24
25
30
25
28
15
23
51.6
58.3
53.3
75
61.7
0.9804
0.8963
0.3382
0.7604
Day 2
Day 3
Day 4
Day 5
Day 6
% dead embryos r =
0.6271 (p=0.2575)
P-value; Chi-square for
trend+
Replicates = 4; n= 302 embryos
+Treatments compared individually with control
www.um.edu.my