Gestational stage affects amniotic epithelial cells phenotype, methylation status, immunomodulatory and stemness properties

Abstract

Stem cells isolated from amniotic epithelium (AECs) have shown great potential in cell-based regenerative therapies. Because of their fetal origin, these cells exhibit elevated proliferation rates and plasticity, as well as, immune tolerance and anti-inflammatory properties. These inherent attitudes make AECs well-suited for both allogenic and xenogenic cellular transplants in animal models. Since in human only at term amnion is easily obtainable after childbirth, limited information are so far available concerning the phenotypic and functional difference between AECs isolated from early and late amnia. To this regard, the sheep animal model offers an undoubted advantage in allowing the easy collection of both types of AECs in large quantity. The aim of this study was to determine the effect of gestational age on ovine AECs (oAECs) phenotype, immunomodulatory properties, global DNA methylation status and pluripotent differentiation ability towards mesodermic and ectodermic lineages. The immunomodulatory property of oAECs in inhibiting lymphocyte proliferation was mainly unaffected by gestational age. Conversely, gestation considerably affected the expression of surface markers, as well the expression and localization of pluripotency markers. In detail, with progression of gestation the mRNA expression of NANOG and SOX2 markers was reduced, while the ones of TERT and OCT4A was unaltered; but at the end of gestation NANOG, SOX2 and TERT proteins mainly localized outside the nuclear compartment. Regarding the differentiation ability, LPL (adipogenic-specific gene) mRNA content significantly increased in oAECs isolated from early amnia, while OCN (osteogenic-specific gene) and NEFM (neurogenic-specific gene) mRNA content significantly increased in oAECs isolated from late amnia, suggesting that gestational stage affected cell plasticity. Finally, the degree of global DNA methylation increased with gestational age. All these results indicate that gestational age is a key factor capable of influencing morphological and functional properties of oAECs, and thus probably affecting the outcome of cell transplantation therapies.

Therapeutic potential of hAECs for early Achilles tendon defect repair through regeneration

“…Moreover, the enhanced development of the inflammatory phase promoted by hAECs could represent another positive event leading to the early tendon healing that mimics what occurs when acute tendinopathies are promptly treated with anti‐inflammatory drugs (Richardson, Dudhia, Clegg, & Smith, ). First of all, as recently demonstrated in vivo for oAECs (Mauro et al, ), hAECs transplantation elicited a lower infiltration of leucocytes, T lymphocytes, and Mφ, thus confirming the in situ persistence of the inherited immunomodulatory activities demonstrated in vitro (Barboni, Russo, Curini, et al, ).…”

Conditioned medium derived from rat amniotic epithelial cells confers protection against inflammation, cancer, and senescence

“…Oct4/Sox2 bind to the proximal region of the Nanog promoter and support its expression. The expression of Nanog is restricted to pluripotent cells, but it is down-regulated upon differentiation in a pattern that is well-conserved in mouse, rat, human [29] and sheep [19]. Here, gene and protein expression of the pluripotency markers confirmed stemness potential of rat AECs.…”

“…Amniotic cells dose-dependently inhibit the proliferation of activated peripheral blood mononuclear cells (PBMCs) [10, 19], but only few studies have focused on the effects of AECs on macrophages [37, 38, 39]. Like CD4 + lymphocytes, the macrophages polarize also toward different phenotypes in response to stimuli.…”

“…The present study was designed to investigate whether rat AECs retain multipotency, plasticity, and immune modulatory properties, and possess anti-proliferative activity against cancer cell lines as described with human [7, 16, 17], equine [18], and ovine [19, 20] AECs. We also investigated whether the conditioned medium (CM) of rat AECs contain soluble factors capable at improving markers of replicative senescence in human fibroblasts.…”

Amniotic Mesenchymal Stromal Cells Exhibit Preferential Osteogenic and Chondrogenic Differentiation and Enhanced Matrix Production Compared With Adipose Mesenchymal Stromal Cells

“…Additionally, enhanced differentiation could have been due in part to the age differences between the younger amnion and older adipose MSCs; donor age and health have been shown to affect MSC properties. 3,4,6,22 …”