Table 1.
Antibodies used in immunohistochemistry.
Figure 1.
Homing of eAT-MSCs (green fluorescence) after transplantation in endometrium of mares with endometrosis.
A) Homing of eAT-MSCs in periglandular space B) eAT-MSC contribution into the whole uterine gland (G). White arrow indicates uterine gland without eAT-MSCs. C) Incorporation of several eAT-MSC (white arrow) into uterine gland epithelia. D–E) eAT-MSCs localization in periglandular space. N-Nucleus. F) Control animals injected with only saline solution. A, B, F = nucleus stained by DAPI (blue). Confocal microscopy: Fluorescence (Fcm) + Digital Interference Contrast (DIC). A–B = Fcm. C–F = Fcm + DIC. Scale bars: A, C, D = 10 µm; B, E, F = 5 µm.
Figure 2.
Vimentin and laminin expression before (at day 0) and after (at days 7, 21 and 60) eAT-MSCs intrauterine transplantation: A–A3 and D–D3 - experimental; B–B3 and D–D3 - control.
A, B) At day 0, vimentin (black arrowheads) localized in damaged epithelia of glands (G) and in fibrotic stromal cells (SC, black arrows). Unaffected epithelia (UE) showed no signs of vimentin expression. A1–A3) At days 7, 21 and 60, the absence of vimentin expression. B1, B2) Vimentin expression is still observed (black arrows) at days 7 and 21, in control. B3) At day 60, control mares showed no signs of vimentin expression. C, D) At day 0, laminin demonstrated high discontinuity of epithelial basal lamina (black arrows) and a diffuse intracytoplasmatic laminin expression in metabolic active fibrotic stromal cells (black arrowheads); C1, C2) At days 7 and 21, unaffected glands (UG) with a diffuse intracytoplasmatic laminin expression were observed. C3) At day 60, the absence of vimentin expression was shown. D1, D2) At days 7 and 21, control maintains same pattern of laminin expression as in (D). D3) At day 60, laminin expression was not visualized in control. Light Microscopy (LM). Scale bars: A–C2 = 50 µm; C3, D2, D3 = 25 µm.
Figure 3.
Smooth-muscle-α-actin (SMA) expression before (at day 0) and after (at day 7) eAT-MSCs intrauterine transplantation.
A, B) At day 0, SMA expression was observed in uterine glands (white arrowhead) and in periglandular fibroblasts (black arrow). A1) At day 7, SMA showed no signs of expression. B) Pattern of SMA expression is similar to A and B. LM. Scale bars: A = 25 µm; A1, B, B1 = 50 µm.
Figure 4.
Cytokeratin 18 (CK18) and Ki-67 expression before (at day 0) and after (at days 7, 21 and 60) eAT-MSCs intrauterine transplantation: A–A3 and D–D3 - experimental group; B–B3 and D–D3 - control.
A, B) At day 0, CK18 (black arrows) localized in damaged epithelia of glands (G). A1–A3) At days 7, 21 and 60, the absence of CK18 expression was observed. B1, B2) At days 7 and 21, CK18 expression was still observed (black arrows) in control. B3) At day 60, control mares showed no signs of CK18 expression. C, D) At day 0, none or a few Ki-67 positive cells were observed. C1, D1) At day 7, amount of Ki-67 positive cells (black arrow) was increased. C2, D2) At day 21, both groups showed positive Ki-67 staining. C3) At day 60, the expression of Ki-67 was still observed. D3) In control, the absence of Ki-67 expression. LM. Scale bars: A–D1 = 50 µm; C2, C3, D2, D3 = 25 µm.
Table 2.
Proliferation rate analyzed by Ki-67 antigen expression.
Figure 5.
Histological analysis of alterations in mares’ endometrium following eAT-MSCs intrauterine transplantation.
A–D3) Mares which received the cells. DE–F3) Control mares. A–F) Morphology of endometrial surface prior eAT-MSCs intrauterine cells transplantation. A1–D1) Day 7 after eAT-MSCs intrauterine transplantation. A2–D2) Same as in (A1–D1) at day 21. A3–D3) Same as in (A1–D1) at day 60. E1–F3) Respective controls. LM. Scale bars: A–F3 = 50 µm.