Journal: Frontiers in Cell and Developmental Biology
Article Title: Evidence for Recipient-Derived Cells in Peribiliary Glands and Biliary Epithelium of the Large Donor Bile Ducts After Liver Transplantation
doi: 10.3389/fcell.2020.00693
Figure Lengend Snippet: Cells that were included in the chimerism counting of the large donor bile ducts showed biliary commitment. (A) Matching procedure to match the fluorescence in situ hybridization (FISH)-treated sections with immunohistochemistry. We used Bioview Duet-3 software to match an image of the FISH-treated sections with immunohistochemistry allowing us to locate high-power FISH photomicrographs within the section and to determine cytokeratin 19 (CK19) expression of individual FISH-treated cells. Two red-colored rectangles from the FISH overview are shown in the CK19 overview in the right image. (B) CK19 immunostainings and FISH photomicrographs of PBG; the two CK19 immunostainings in the left images correspond with the two red-colored rectangles in the CK19 overview in panel A. CK19 is a marker for biliary epithelial cells. Images that are horizontal aligned show the same area in the cross-section. Arrowheads in the left image point toward CK19+ cells, these same cells are pointed out in the FISH photomicrograph and the merged image on the right side. These CK19+ epithelial cells were included in the chimerism counting. (C) Immunohistochemistry for cluster of differentiation 45 (CD45) with matching FISH photomicrographs. CD45 is a marker for inflammatory cells. Images that are horizontal aligned show the same area in the cross-section. Arrowheads point toward CD45+ cells in the CD45 immunostaining, FISH photomicrograph, and the merged image. (D) Chimerism counting in CD45 and CK19 immunostainings merged with FISH photomicrographs. Black arrows show that the left and right image in panel D are magnifications of the upper CD45-FISH merged image in panel C and the lower CK19-FISH merged image in panel B, respectively. Male recipient-derived cells, encircled by a blue dotted line, present with red and green spots indicating Y and X chromosomes, respectively. Female donor-derived cells, encircled by a pink dotted line, present with two green dots indicating two X chromosomes. In the image on the left side, a CD45 negative PBG cell is depicted by an arrowhead (see also corresponding images in panel C ) and magnified in the yellow box in the lower right corner. This recipient-derived cell can therefore be classified as a non-inflammatory PBG cell (i.e., cholangiocyte). In the image on the right side, a CK19+ epithelial cell is pointed out by an arrowhead and magnified in the yellow box in the lower right corner. Cells that are encircled by a blue or pink dotted line were included in the chimerism counting. (B–D) Original magnification ×60.
Article Snippet: Fluorescence microscopy was performed using an Olympus BX63 microscope (Olympus corporation, Leiderdorp, Netherlands) equipped with a CCD video camera and Bioview duet-3 Software.
Techniques: Fluorescence, In Situ Hybridization, Immunohistochemistry, Software, Expressing, Marker, Immunostaining, Derivative Assay
