Abstract
The secretion of potential therapeutic factors by mesenchymal stem cells (MSCs) has aroused much interest given the benefits that it can bring in the field of regenerative medicine. Indeed, the in vitro multipotency of these cells and the secretive capacity of both angiogenic and immunomodulatory factors suggest a role in tissue repair and regeneration. However, during culture, MSCs rapidly lose the expression of key transcription factors associated with multipotency and self-renewal, as well as the ability to produce functional paracrine factors. In our study, we show that a three-dimensional (3D) culture method is effective to induce MSC spheroid formation, to maintain the multipotency and to improve the paracrine activity of a specific population of human amnion-derived MSCs (hAMSCs). The regenerative potential of both 3D culture-derived conditioned medium (3D CM) and their exosomes (EXO) was assessed against 2D culture products. In particular, tubulogenesis assays revealed increased capillary maturation in the presence of 3D CM compared with both 2D CM and 2D EXO. Furthermore, 3D CM had a greater effect on inhibition of PBMC proliferation than both 2D CM and 2D EXO. To support this data, hAMSC spheroids kept in our 3D culture system remained viable and multipotent and secreted considerable amounts of both angiogenic and immunosuppressive factors, which were detected at lower levels in 2D cultures. This work reveals the placenta as an important source of MSCs that can be used for eventual clinical applications as cell-free therapies.
Highlights
Adult stem cells are extensively used for regenerative medicine because of their multilineage potential and regenerative properties
We established 90% of human amnion-derived MSCs (hAMSCs) culture from 10 processed placentas (Figure 2(a)), and from these cells grown in a suspended state (3D), we observed the generation of spheroids (Figure 2(b))
It was observed that the human mesenchymal stem cells (MSCs) markers CD90, CD73, and CD13 were expressed in all cultures, whereas the cells were negative for CD45 and HLA-DR
Summary
Adult stem cells are extensively used for regenerative medicine because of their multilineage potential and regenerative properties. These cells exist in different tissues, including fat [1], bone marrow [2], the umbilical cord [3], and placenta tissue [4], where they participate in the maintenance of stem cell niches and tissue homoeostasis [5]. Though the pathophysiologic functions of mesenchymal stem cells (MSCs) are under investigation, the in vitro multipotency of these cells suggests a role in tissue regeneration, wound healing, and/or tissue repair after transplantation [6]. MSCs are capable of self-renewal and differentiation into several mesenchymal lineages both in vitro and in vivo, including fat, bone, and cartilage [7]. Despite the availability of various cell sources for the use of MSCs in the field of regenerative medicine, the ethical
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