Corradetti Bruna, Correani Alessio, Romaldini Alessio, Marini Maria Giovanna, Bizzaro Davide, Perrini Claudia, Cremonesi Fausto, Lange-Consiglio Anna
Department of Life and Environmental Sciences, Università Politecnica delle Marche, Ancona, Italy.
Large Animal Hospital, Reproduction Unit, Università degli Studi di Milano, Lodi, Italy.
PLoS One. 2014 Oct 31;9(10):e111324. doi: 10.1371/journal.pone.0111324. eCollection 2014.
Amniotic membrane-derived mesenchymal cells (AMCs) are considered suitable candidates for a variety of cell-based applications. In view of cell therapy application in uterine pathologies, we studied AMCs in comparison to cells isolated from the endometrium of mares at diestrus (EDCs) being the endometrium during diestrus and early pregnancy similar from a hormonal standpoint. In particular, we demonstrated that amnion tissue fragments (AM) shares the same transcriptional profile with endometrial tissue fragments (ED), expressing genes involved in early pregnancy (AbdB-like Hoxa genes), pre-implantation conceptus development (Erα, PR, PGRMC1 and mPR) and their regulators (Wnt7a, Wnt4a). Soon after the isolation, only AMCs express Wnt4a and Wnt7a. Interestingly, the expression levels of prostaglandin-endoperoxide synthase 2 (PTGS2) were found greater in AM and AMCs than their endometrial counterparts thus confirming the role of AMCs as mediators of inflammation. The expression of nuclear progesterone receptor (PR), membrane-bound intracellular progesterone receptor component 1 (PGRMC1) and membrane-bound intracellular progesterone receptor (mPR), known to lead to improved endometrial receptivity, was maintained in AMCs over 5 passages in vitro when the media was supplemented with progesterone. To further explore the potential of AMCs in endometrial regeneration, their capacity to support resident cell proliferation was assessed by co-culturing them with EDCs in a transwell system or culturing in the presence of AMC-conditioned medium (AMC-CM). A significant increase in EDC proliferation rate exhibited the crucial role of soluble factors as mediators of stem cells action. The present investigation revealed that AMCs, as well as their derived conditioned media, have the potential to improve endometrial cell replenishment when low proliferation is associated to pregnancy failure. These findings make AMCs suitable candidates for the treatment of endometrosis in mares.
羊膜来源的间充质细胞（AMCs）被认为是各种基于细胞的应用的合适候选者。鉴于细胞疗法在子宫疾病中的应用，我们将AMCs与从处于发情间期的母马子宫内膜分离的细胞（EDCs）进行了比较研究，从激素角度来看，发情间期和妊娠早期的子宫内膜相似。特别是，我们证明羊膜组织碎片（AM）与子宫内膜组织碎片（ED）具有相同的转录谱，表达参与早期妊娠的基因（AbdB样Hoxa基因）、植入前胚胎发育（Erα、PR、PGRMC1和mPR）及其调节因子（Wnt7a、Wnt4a）。分离后不久，只有AMCs表达Wnt4a和Wnt7a。有趣的是，发现前列腺素内过氧化物合酶2（PTGS2）在AM和AMCs中的表达水平高于其子宫内膜对应物，从而证实了AMCs作为炎症介质的作用。已知能提高子宫内膜容受性的核孕酮受体（PR）、膜结合细胞内孕酮受体成分1（PGRMC1）和膜结合细胞内孕酮受体（mPR）的表达，在体外培养基中添加孕酮的情况下，AMCs传代5次后仍能维持。为了进一步探索AMCs在子宫内膜再生中的潜力，通过在transwell系统中将它们与EDCs共培养或在AMC条件培养基（AMC-CM）存在下培养，评估它们支持驻留细胞增殖的能力。EDC增殖率的显著增加表明可溶性因子作为干细胞作用介质的关键作用。本研究表明，当低增殖与妊娠失败相关时，AMCs及其衍生的条件培养基有潜力改善子宫内膜细胞补充。这些发现使AMCs成为治疗母马子宫内膜异位症的合适候选者。
