epithelial origin
isolated BOECs
confirmed
subjecting
immunocytochemical
staining
expression
cytokeratins
using
monoclonal
mouse
antihuman
cytokeratin
antibodies
biotinylated goat
antimouse
streptavidin–
horseradish
peroxidase
substrate-chromogen
according
manufacturer
instructions.
Bovine
oviductal
epithelial cells
four-well
culture plates
freshly retrieved BOECs
first-passage BOECs
culture
period
flask
trypsinized
reseeded
glass inlets, and (3) as freshly retrieved BOECs
cultured as monolayers on hanging inserts in a PCC system.
Cytokeratin-stained cells were compared to nonstained
cells, and a fibroblast culture of fetal bovine cartilage was
used as a negative control, and nonspecific bindingreactions of the secondary and tertiary antibodies were
excluded. Morphologic evaluation was performed by an
inverted light microscope, and the imaging software of
CellD* imaging was used to assess the ratio of cytokeratinpositive
and cytokeratin-negative cells.