Publication: In vitro modifications in the proliferation
Authors
Carretero, J. ; Rubio, M. ; Navarro, N. ; Prieto, P. ; Vázquez, R.J. ; Sanchez, F. ; Vazquez, R.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
In order to establish the correlation between
in vitro proliferation rate and morphometric variations of
prolactin immunoreactive cells, a morphometric study
was carried out in rat pituitary monolayer cultures by
means of the double immunocytochemical staining
methods employing mouse monoclonal antiproliferative
cell nuclear antigen (PCNA) and rabbit anti-prolactin
(PRL) as primary antibodies. PCNA was found to be an
adequeate marker for proliferation in pituitary
monolayer cultures. 48.35+2.78% of the cells present in
the culture were in active cell cycle after 3 days of
incubation and a similar proportion, 54.93&2.83% was
found after 7 days. On the 3rd day, PRL imrnunopositive
cells accounted for 15.16*0.21% of the total cellular
content in the dishes and 8.68*0.12% of the PCNA
immunoreactive cells were also PRL immunopositive
cells and, 60.95112.65% of PRL cells stained for PRL
and PCNA. On the 7th day, an increase to 32.18f 0.60%
of PRL cells was found; the PCNA and PRL cells
accounted for 60.32*2.34% of the total PRL cells, and
19.88I1.09% of the PCNA reactive cells stained for
PRL. Additionally, the morphometric analysis performed
after 3 or 7 days of incubation showed that, while the
size of PRL cells remained unmodified, the nuclear area
had increased on the 7th day in relation to the 3rd day
(pc0.01). These results suggest: 1) PCNA is a valid
proliferative marker for pituitary cells in cultures; 2) a
very high percentage of the PRL cells was in early
proliferation; 3) on the 7th day of incubation, the
proliferative rate of PRL cells was very similar to that
observed on the 3rd day, suggesting a maintained
proliferation for PRL cells at early incubation phases;
and 4) the cellular activity, expressed as variations in the
nuclear size, was higher on the 7th day than on the 3rd
day; in addition, the numerical density of PRL cells
increased.
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