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Publication of the International Union Against Cancer
Publication de l’Union Internationale Contre le Cancer
Int. J. Cancer: 71, 310 (1997)
r 1997 Wiley-Liss, Inc.
LETTER TO THE EDITOR
Dear Sir,
Identification of nude mice in tumorigenicity assays
A test widely used to determine the effects of gene alterations
on cell malignancy is implantation in vivo and monitoring of
tumor formation. To perform such experiments with human
cells, an immunocompromised animal host is required. Perhaps
the most commonly used animal system for such tumorigenicity
assays is the athymic ‘‘nude’’ mouse (Giovanella et al., 1974).
Although this system is limited by false-negative results, it is
widely accepted as an indicator of malignancy. Take rates can
be improved by irradiation (30–60 Gy) or tumor cell implantation in Matrigel (Pretlow et al., 1991). One practical issue in
dealing with these animals is precise identification of each
individual mouse. Current practices on a small scale include
the use of pen marks on tails, ear piercing or toe cutting. The
shortcomings of these are loss of pen marks due to grooming
and infringing on animal integrity by piercing or cutting. Here,
I propose the use of a simple alternative method to label ‘‘nude’’
animals—namely, injection of colloidal carbone. Briefly, 5–10
µl of colloidal carbon suspension (China Ink, Pelikan, #211862)
sterilized by autoclaving is injected s.c. with a 25G needle
above the thigh of the animal (Fig. 1). Even smaller marks can
be obtained by depositing a drop of ink on the skin and simply
piercing through the drop with an empty needle. The ink easily
penetrates the skin through capillary force. These marks are
indelible, do not harm the animal and do not produce side
effects such as bleeding or infections (over 500 mice tested so
far). Usually, 4 or 5 animals are kept per cage, so a numbering
system from 1 to 5 is suffıcient. This can be achieved with
maximum of 2 injections per animal: one and two spots on the
left hind leg (for #1 and #2), or on the right hind leg (for #3 and
FIGURE 1 – Nude mouse #4 injected twice s.c. in the right hind leg
with 5 µl of sterilized colloidal carbon suspension.
#4), and one spot on each side for #5. Therefore, I recommend
the use of this simple tattooing procedure for the marking of
‘‘nude’’ animals for small-scale tumorigenicity experiments.
Yours sincerely,
Erwin G. VAN MEIR
Laboratory of Tumor Biology and Genetics, Neurosurgery
Department, University Hospital (CHUV), 1011 Lausanne, Switzerland
Received 5 November 1996
REFERENCES
GIOVANELLA, B.C., STEHLIN, J.S. and WILLIAMS, L.J., Heterotransplantation
of human malignant tumors in ‘‘nude’’ thymusless mice. II. Malignant
tumors induced by injection of cell cultures derived from human solid
tumors. J. nat. Cancer Inst., 52, 921–930 (1974).
PRETLOW, T.G., DELMORO, C.M., DILLEY, G.G., SPADAFORA, C.G. and
PRETLOW, T.P., Transplantation of human prostatic carcinoma into nude
mice in Matrigel. Cancer Res., 51, 3814–3817 (1991).
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