Amniotic Membrane-Derived Mesenchymal Cells and Their Conditioned Media: Potential Candidates for Uterine Regenerative Therapy in the Horse

Abstract

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-implantantion 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.

Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in endometrial cells in an in-vitro model

“…A detailed characterization of these cells was performed in the paper of Corradetti et al [39]. In this study, a molecular characterization of EDCs was performed only at passage (P)0 as a de facto control for gene expression.…”

“…After isolation from endometrial tissue, cells were analyzed to detect the expression of specific endometrial genes as previously reported [39]. Total RNA was extracted from endometrial cells immediately after isolation (P0) using TRI Reagent Solution (Life Technologies, Monza, Italy) and conventional RT-PCR was performed with RBC Taq DNA Polymerase (RBC Bioscience) using previously optimized primers [39].…”

“…Total RNA was extracted from endometrial cells immediately after isolation (P0) using TRI Reagent Solution (Life Technologies, Monza, Italy) and conventional RT-PCR was performed with RBC Taq DNA Polymerase (RBC Bioscience) using previously optimized primers [39]. The primer sequences and conditions are shown in Tables 1 and 2.…”

Enhanced osteogenic potential of mesenchymal stem cells from cortical bone: a comparative analysis

“…Proliferation rate was determined in triplicate on AD-, BM-, and CBF-MSCs as previously reported [46,47]. Doubling time was assessed from P1 to P10 in normoxic and hypoxic conditions to determine the effect of lowtension oxygen on cell proliferation.…”

Nanotechnology for mesenchymal stem cell therapies

“…They have been proven effective in endometrial cell replenishment when low proliferation is associated to pregnancy failure in vitro [24] and to be sufficient to stimulate the structural and functional regeneration of cardiac [25,26], renal [27,28], tendon [29], and spinal cord [30] tissues Figure 1 summarizes the MSC features that make them a powerful tool for therapeutic purposes.…”