Fig 1.
The different steps of the surgical protocol.
D-10: Synchronization of the estrous cycle by an intra-vaginal progesterone sponge D0: Day of surgery; T: Time in minutes.
Fig 2.
Photographs showing various steps of uterus auto-transplantation.
a) Dissection between uterine vessels and ureter; (b) Excised uterus on the backtable; (c) anastomosis; (d) uterine anastomosis at dissection. 1 = uterine artery; 2 = ureter; 3 = umbilical artery; 4 = utero-ovarian vein; 5 = external iliac artery; 6 = external iliac vein.
Fig 3.
Histology of the upper part of a uterine horn in sheep.
(a) Hematoxylin–eosin stained section of the upper horn. Arrows delineate the various tissues of the uterine wall (X20 magnification used for stereology). (b) Hematoxylin-eosin stained section of the upper horn (X40). Arrows indicate infiltrating neutrophils. S, Stroma; G, endometrial Gland; A, Artery; M, Muscle; MCT, Muscular Connective Tissues; LE, Luminal Epithelium and OE, Oedema.
Table 1.
Primer sequences.
Fig 4.
Time course of pH, gas pressure and lactates in venous samples of transplanted uterus.
Blood was collected at various time points during the 180 min perfusion time. a) pH; b) pCO2/pO2; c) lactate. Dotted lines indicate the maximum and minimum range in samples taken before harvesting the graft. Individual values (rounds) and medians (line) are shown.
Fig 5.
Neutrophils density in the endometrium of auto-transplanted uterus.
Median density +/- SEM is presented for each phenotype (control group, group 1: Reperfusion of 2 veins, group 2: Reperfusion of 1 vein). For every ewe, median density of upper and lower part of the horns was calculated.
Table 2.
Stereology: Histological structures (volume %) comparison of medians [1st Quartile-3rd Quartile] of upper horns between Group 1 (2 veins) Group 2 (1 vein) and control group.
Fig 6.
Expression levels of a selection of genes in the endometrium of ovine auto-transplanted uteri.
(a) Selection of genes involved in inflammation, apoptosis, (b) selection of immune-related genes. For each group, mean expression and standard error of the mean (SEM) were presented as histograms. No surgery of the uterine vascularization was carried out in the control group (3 ewes). In groups 1 (3 ewes) and 2 (4 ewes), uterine horns were identified according to their vascularization; A = artery; V vein; 1 = reperfusion; 0 = absence of reperfusion. *, p<0.05; **, p<0.01, ***, p<0.001. AU: Arbitrary unit.
Fig 7.
Quantification of PTGS2 protein by Western Blot in the endometrium of auto-transplanted ovine uteri.
The amount of PTGS2/protein was normalized to that of actin B protein (ACTB). For each group (control, group 1 and group 2), three ewes were analyzed. A = artery; V vein; 1 = reperfusion; 0 = absence of reperfusion. S: PTGS2 Standart.
Fig 8.
Immunolocalization of PTGS2 protein in the endometrium of auto-transplanted ovine uteri.
Immunohistochemistry was performed using sections cut from ovine endometrium. For each uterus, the two horns are labelled horn A and horn B to make the interpretation easier. Uterine vascularization is indicated in the top left corner of each microphotograph. CTR: Control; A = artery; V vein; 1 = reperfusion; 0 = absence of reperfusion. No surgery of the uterine vascularization was carried out in the control group. Specificity of ovine PTGS2 labelling was carried out using (i) a section of ovine endometrium sampled from a ewe at day 14 of gestation (14DG), and (ii) a section of endometrium sampled from a control uterus incubated in the absence of the primary antibody (No PTGS2 AB). Bars = 1250 μm (insert: 250 μm).