Neural differentiation of human adipose‐derived mesenchymal stem cells induced by glial cell conditioned media

Abstract

Adipose-derived mesenchymal stem cells (ASCs) may transdifferentiate into cells belonging to mesodermal, endodermal, and ectodermal lineages. The aim of this study was to verify whether a neural differentiation of ASCs could be induced by a conditioned medium (CM) obtained from cultures of olfactory ensheathing cells (OECs) or Schwann cells (SCs). ASCs were isolated from the stromal vascular fraction of adipose tissue and expanded for 2-3 passages. They were then cultured in OEC-CM or SC-CM for 24 hr or 7 days. At each stage, the cells were tested by immunocytochemistry and flow cytometer analysis to evaluate the expression of typical neural markers such as Nestin, PGP 9.5, MAP2, Synapsin I, and GFAP. Results show that both conditioned media induced similar positive effects, as all tested markers were overexpressed, especially at day 7. Overall, an evident trend toward neuronal or glial differentiation was not clearly detectable in many cases. Nevertheless, a higher tendency toward a neuronal phenotype was recognized for OEC-CM (considering MAP2 increases). On the other hand, SC-CM would be responsible for a more marked glial induction (considering GFAP increases). These findings confirm that environmental features can induce ASCs toward a neural differentiation, either as neuronal or glial elements. Rather than supplementing the culture medium by adding chemical agents, a "more physiological" condition was obtained here by means of soluble factors (cytokines/growth factors) likely released by glial cells. This culture strategy may provide valuable information in the development of cell-based therapeutic approaches for pathologies affecting the central/peripheral nervous system.

Conditioned Media From Glial Cells Promote a Neural-Like Connexin Expression in Human Adipose-Derived Mesenchymal Stem Cells

“…In accordance with previous studies ( Calabrese et al, 2015 ), cells were immunopositive for typical MSC markers (CD 44, CD 73, CD 90, and CD 105) and immunonegative for typical hematopoietic stem cell markers (CD 14, CD 34, and CD 45). Moreover, as reported recently ( Lo Furno et al, 2018 ), an ASC neural-like phenotype by using OEC-CM or SC-CM was here confirmed by the increased immunopositivity for Nestin, PGP9.5, and GFAP (Figure 1 ).…”

“…In our previous studies ( Lo Furno et al, 2013b , 2018 ), a successful differentiation of ASCs toward neural-like elements was achieved by using a conditioned medium (CM) from cultures of Schwann cells (SCs) or olfactory ensheathing cells (OECs). This differentiation was documented by the increased expression of typical neural markers (Nestin, PGP9.5, MAP2, Synapsin I, GFAP).…”

Neural differentiation of canine mesenchymal stem cells/multipotent mesenchymal stromal cells

“…Neurospheres generated from the canine ASCs were used to enrich for neural lineages expanded enough to transplant (19) and when grown in hypoxia showed higher expression of neuronal marker nestin (20). Recently, neural differentiation of human ASCs was evoked by conditioned medium obtained from glial cells (21).…”

Therapeutic Effects of Genetically Modified Wharton’s Jelly Mesenchymal Stem Cells Expressing Erythropoietin on Cancer-related Anemia in Mice Model

“…Mesenchymal stem cells (MSCs) as desirable cell carriers can be easily obtained, expanded, and genetically engineered to express and secret therapeutic proteins in-vivo (21)(22)(23)(24). Human Wharton's jelly mesenchymal stem cells (hWJMSCs) are multipotent stem cells which showed the potential to differentiate into mesodermal, ectodermal and endodermal lineages (25,26). Compared to adult and fetal stem cells, hWJMSCs show a higher proliferation rate and minimum stimulation of immune and in ammatory systems (27)(28)(29).…”