Biology Reference
In-Depth Information
between mice and humans do exist, mostly based on the type of
sequestered cells in brain microvasculature. However, since there
is no alternative to study severe malaria and to develop new con-
cepts of treatment, the murine model of cerebral malaria provides
experimental evidences that need to be confirmed in humans (see
Note 3).
2
Materials
Animal experiments are conducted according to the Institutional
Animal Care and Use Committee guidelines and in agreement
with international rules. The model used in the lab is CBA/J
mice 6 weeks old inoculated with 10
6
Plasmodium berghei ANKA
(see Note 4).
2.1 Inoculum:
Plasmodium berghei
ANKA Parasitized Red
Blood Cells
1.
P. berghei
ANKA aliquots of 1 mL (concentration = 10
7
GRP/
mL) are stored in liquid nitrogen. Frozen aliquots must be
discarded after 1 year.
2. Preheat sterile phosphate buffered saline (PBS) at 37°C.
3. Remove aliquots of
P. berghei
ANKA from liquid nitrogen and
proceed to a quick defrost of the aliquot at 37°C.
4. Slowly add 1 mL of sterile PBS to 1 mL aliquot of
P. berghei
ANKA and homogenize the suspension carefully.
5. Start intraperitoneal (IP) infection less than 5 min after inocu-
lum preparation.
2.2 Mice: CBA/J
( See Note 5 )
Six-week-old female CBA/J mice weighting 18-20 g are used.
Allow mice to adapt to their environment for 7 days before experi-
ments. Choose the number of mice needed per group according to
the expected effect and statistical analysis (see Note 6).
2.3
Drugs
1. Erythropoietin beta (see Notes 7 and 8): Dilute erythropoietin
beta in sterile saline to obtain a final concentration of 100 IU/
mL. After dilution, Epo is stable for 2 h. Calculate the volume
of Epo according to each mouse weight. For a mouse weight-
ing 20 g and an expected posology of 1,000 IU/kg, inject
200
L of EPO 100 IU/mL using intraperitoneal route.
2. Artesunate (Molecular weight: 384.42; Molecular formula:
C
19
H
28
O
8
): Dilute 8 mg of artesunate powder in 133
μ
μ
L of
sodium bicarbonate 4.2%. Then, add 877
L of sterile saline
to obtain 8.0 g/L concentration. Calculate the injected vol-
ume of artesunate solution according to each mouse weight.
For a mouse weighting 20 g and an expected posology of
40 mg/kg, inject 100
μ
μ
L of artesunate 8.0 g/L using intrap-
eritoneal route.
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