Cholinergic control of cell growth and migration in undifferentiated and schwann cell–like differentiated adipose-mesenchimal stem cells.

Peripheral nerve injury is a common disease and represents a major economic burden for society. The development of novel strategies, including cell therapy, to enhance peripheral nerve regeneration is therefore of great social and clinical relevance. The use of synthetic nerve conduits, in combination with different cell types may represent a promising therapy. The great regenerative capacity of peripheral nervous system (PNS) is due to a permissive environment provided by Schwann cells that proliferate, migrate and release growth factors either during development or after nerve lesions. Mesenchimal stem cells (MSCs) are an attractive cell source for nerve tissue regeneration. They are able to self-renew and possess multi potent differentiation properties. In particular adipose MSCs (A-MSCs) appear the most promising source of MSCs. Recent studies have shown that A-MSCs can be differentiated in Schwann-like cells, representing an alternative and reliable source of peripheral glial cells. Acetylcholine (ACh), the main neurotransmitter in central and PNS, has the property to modulate neurite outgrowth and to control Schwann cell proliferation and differentiation. ACh plays important role also in non-neural tissue, but its functions in mesenchymal stem cells (MSC) has been poorly investigated. In present work we have characterized the muscarinic cholinergic agonist effects in rat A-MSC and in differentiated Schwann-like obtained from A-MSCs. Analysis by RT-PCR has demonstrated that the A-MSCs express all muscarinic receptor subtypes. MTT analysis and wound healing assay have also demonstrated that the selective activation of M2 receptors caused an inhibition of cell growth and migration of MSCs indicating the ACh as possible modulator of MSC proliferation and migration. In Schwann cell-like derived from A-MSC, similarly to that observed in Schwann cells, the M2 muscarinic agonist caused a decrease of cell proliferation without affecting cell survival. Further analysis are addressed to evaluate the capability of these receptors to mediate the differentiative processes in Schwann cell-like, as previously observed in Schwann cells. In conclusion, we hypothesize that a combination of autologous MSC, differentiated in Schwann-like, and selective ACh mimetics may represent a successful strategy to achieve better results in peripheral nerve regeneration