Horse mesenchymal stem cells (MSC) are potential anti-inflammatory tools for post-breeding induced endometritis (PBIE). In this research MSCs isolated from the endometrium or subcutaneous fat of the same donors were infused iu into mares with PBIE for assessment of their anti-inflammatory action and engraftment. PBIE was induced in nine gynecologically healthy mares by iu infusion of 500 million dead sperm in saline. Inflammatory markers were analyzed in uterine lavages and biopsies immediately before (phase I) and 3 h after infusion of sperm (phase II). Measurements: polymorph nuclear cells (PMN), proteins IL-6 and TNFα (ELISA in the lavages) and immunostaining in biopsies, transcripts of IL-1α, 6, 8, 10, TNFα and COX2 (qPCR of pelleted lavages). At 24 h after sperm deposition (phase III), mares were instilled iu with 20 ml of saline containing 2 × 107 adipose MSCs (n = 3, group 1) or endometrial MSCs (n = 3, group 2). Cells were labeled previously with carboxyfluorescein diacetate succinimidyl ester (CFDA SE). A third group (n = 3) received 20 mL of sterile saline alone. After 48 h another biopsy/lavage were done and the same parameters analyzed. For engraftment, additional biopsies were taken at days 10 and 30 of sperm infusion and analyzed by confocal microscopy. Dead sperm in saline markedly increased PMNs counts, IL-6 and TNFα expression in the ELISA (p < 0.05) and immunostaining. In phase III a significant reduction (p < 0.0001) of PMN was found in all samples, including control mares. A decrease (p < 0.05) of IL-6 and TNF-α was detected by ELISA, in the groups that received MSC, but not in control group. In the aMSC-treated group, a significant decrease was found in the expression of (IL1α, p = 0.0003; IL-6 p 0.04; IL-8, p = 0.006, TNFα p = 0.004). Expression of IL-10 and COX2 remained unchanged (p = 0.08). In the mares that received eMSC, IL-6 and 8 decreased significantly (p = 0.01), IL-10 increased (p = 0.009), while TNFα, COX2 and IL1α did not significantly change their expression. In the engraftment experiment CFDA label was found sparingly in all the samples analyzed until day 30, mainly at the stromal compartment of the endometrium. No differences in the engraftment pattern was found among cell origins. We conclude that inoculation of MSCs significantly reduced inflammation independently of the origin of the cells and that cells perform limited engraftment detectable after one month of infusion. These findings can be of help for the design of new anti-inflammatory therapies of uterine diseases in mares.

中文翻译：

---

评估马子宫内膜和脂肪间充质干细胞在繁殖后诱发子宫内膜炎的体内模型中的抗炎和植入潜力

马间充质干细胞 (MSC) 是育种后诱发子宫内膜炎 (PBIE) 的潜在抗炎工具。在这项研究中，从相同供体的子宫内膜或皮下脂肪中分离出的 MSC 被注入含有 PBIE 的母马，以评估它们的抗炎作用和植入。通过将 5 亿个死精子在生理盐水中进行 iu 输注，在 9 匹妇科健康的母马中诱导了 PBIE。在输注精子之前（I 期）和输注后 3 小时（II 期）立即在子宫灌洗和活检中分析炎症标志物。测量：多形核细胞 (PMN)、蛋白质 IL-6 和 TNFα（灌洗液中的 ELISA）和活检中的免疫染色、IL-1α、6、8、10、TNFα 和 COX2 的转录物（沉淀灌洗液的 qPCR）。在精子沉积后 24 小时（III 期），给母马注射 20 ml 含 2 × 107 脂肪 MSC（n = 3，第 1 组）或子宫内膜 MSC（n = 3，第 2 组）的盐水。细胞预先用羧基荧光素二乙酸琥珀酰亚胺酯 (CFDA SE) 标记。第三组 (n = 3) 仅接受 20 mL 无菌盐水。48 小时后进行另一次活检/灌洗并分析相同的参数。对于植入，在精子输注的第 10 天和第 30 天进行额外的活组织检查，并通过共聚焦显微镜进行分析。盐水中的死精子显着增加了 ELISA 中的 PMN 计数、IL-6 和 TNFα 表达（p < 0.05）和免疫染色。在阶段 III 中，所有样本（包括对照母马）的 PMN 均显着减少（p < 0.0001）。ELISA 检测到接受 MSC 的组中 IL-6 和 TNF-α 的减少（p < 0.05），而对照组没有。在aMSC处理组中，发现（IL1α，p = 0.0003；IL-6 p = 0.04；IL-8，p = 0.006，TNFα p = 0.004）的表达显着降低。IL-10 和 COX2 的表达保持不变（p = 0.08）。在接受 eMSC 的母马中，IL-6 和 8 显着减少（p = 0.01），IL-10 增加（p = 0.009），而 TNFα、COX2 和 IL1α 没有显着改变它们的表达。在移植实验中，CFDA 标签很少出现在所有分析的样品中，直到第 30 天，主要是在子宫内膜的基质隔室中。在细胞起源中没有发现植入模式的差异。我们得出的结论是，MSC 的接种显着减少了炎症，而与细胞的起源无关，并且细胞在输注 1 个月后进行可检测到的有限植入。