%0 Journal Article
%T Peripheral Blood Lymphocyte Culture of Rhinella arenarum
%A R. Ag¨ąero
%A S. M. Marsa
%A L. E. Moreno
%A M. E. Vasquez Gomez
%J Open Access Library Journal
%V 4
%N 12
%P 1-8
%@ 2333-9721
%D 2017
%I Open Access Library
%R 10.4236/oalib.1104212
%X
The cytogenetic identification is important for the
characterization of an organism. In amphibians, the direct method is a
technique of routine for chromosomal characterization, but is necessary for the sacrifice of the copies. The aim of this work is
to develop the technique of lymphocyte cultures for the
species Rhinella arenarum without sacrifice of specimen. Materials and Methods:
Male specimensˇŻ Rhinella arenarum were collected at San Luis; the blood sample was obtained
by cardiac puncture. The mediums tested were: the culture media: RPMI 1690 with
HEPES and Glutamine, MEM and F10; and different volumes of: phytohemagglutinin,
penicillin-streptomycin, fetal bovine serum and colchicine. Results: For the
standardization of the protocol we rely on cell culture techniques of fish and
human. The following parameters were standardized: volume blood: volume of 100
¦Ěl; culture medium was chosen: the best results were observed with RPMI 1640;
fetal bovine serum: he worked with a volume of 500 ¦Ěl in the case of RPMI 1640
and F10, for MEM 1000 ¦Ěl was used; phytohemagglutinin: it was observed that
large volumes of this reagent agglutinated cells, and hence the greater number of metaphases was obtained
with 10 ¦Ěl; colchicine: best chromosome size was observed with 100 ¦Ěl and an incubation time of 12 h. Conclusion: The standardized protocol is a simple
and inexpensive technique that does not require equipment or facilities of high
complexity and specimen are kept alive.
%K Amphibian
%K Cytogenetics
%K Metaphase
%U http://www.oalib.com/paper/5291922