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Abstract
We developed a novel technique to detect S-phase skin cells by applying bromodeoxyuridine
(BrdU) epicutaneously using an occlusive dressing (OD) method. BrdU was scarcely absorbed
from the skin with a simple epicutaneous application, whereas the incorporation of
BrdU was very well promoted with the use of our OD method. We applied BrdU on the
backs of rats using this method and investigated the conditions required for an optimal
response, with a special focus on the period of application, the concentration of
BrdU used and vehicles suitable for the immunocytochemical staining of this agent.
From these experiments, we were able to detemine that an application time of at least
60 min was necessary to label S-phase cells, a 2% concentration of BrdU was needed
to obtain consistent labeling and aqueous vehicles are satisfactory solvents for BrdU
preparations. Epidermal keratinocytes and S-phase cells in the upper portion of dermis
were clearly labeled after either intraperitoneal injection of BrdU or after administration
by means of the OD method. To ascertain whether this latter method could provide an
effective alternative to intraperitoneal injection, we compared the labeling patterns
of both methods with respect to the speed of migration of BrdU-labeled basal cells
from the basal layer to the horny layer of epidermis. Using either of these two methods,
basal keratinocytes were labeled immediately after administration. Three days after
the first administration, BrdU-labeled cells were detected in the middle layer of
the epidermis, but after 8 days, they were no longer evident in epidermal tissue.
As another means of comparing both methods, we used antibody to proliferating cell
nuclear antigen (PCNA) and compared the ratio of PCNA-positive basal cells to BrdU-labeled
basal cells. The number of PCNA-positive cells was about 4.6 times greater than the
number of BrdU-labeled basal cells by both methods. We concluded that the OD method
could be used as a substitute for intraperitoneal injection in order to observe cell
kinetics using bromodeoxyuridine.
Keywords
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Article info
Publication history
Accepted:
April 26,
1996
Received in revised form:
April 15,
1996
Received:
February 5,
1996
Identification
Copyright
© 1997 Published by Elsevier Inc.
