Carbachol, along with calcium, indicates new strategy in neural differentiation of human adipose tissue-derived mesenchymal stem cells in vitro.

Content Provider	Europe PMC
Author	Masoumi, Niloofar Ghollasi, Marzieh Eftekhari, Elahe Ghiasi, Mohsen Raheleh Halabian
Copyright Year	2023
Abstract	IntroductionOver the past few years, stem cells have represented a promising treatment in neurological disorders due to the well-defined characteristics of their capability to proliferate and differentiate into any cell type, both in vitro and in vivo. Additionally, previous studies have shown that calcium signaling modulates the proliferation and differentiation of neural progenitor cells. The present study investigated the effect of carbachol (CCh), a cholinergic agonist activating acetylcholine receptors, with and without calcium, on the neural differentiation of human adipose tissue-derived mesenchymal stem cells (hADSCs) in neural media, including forskolin and 3-isobutyl-1-methyl-xanthine and retinoic acid.MethodsFor this purpose, first, the MTT assay and acridine orange staining were studied to obtain the optimal concentration of CCh. Next, the differentiation tests, such as cellular calcium assay as well as evaluation of qualitative and quantitative expression of neuronal index markers through immunofluorescence staining and gene expression analysis, respectively, were performed on days 7 and 14 of the differentiation period.ResultsAccording to the results, CCh at 1 μM concentration had no cytotoxicity on hADSCs and also induced cell proliferation. Furthermore, CCh with and without calcium increased the expression of neural-specific genes (NSE, MAP2, β–III–tubulin, and MAPK3) and proteins (γ-enolase, MAP2, and β–III–tubulin) as well as the amount of calcium in differentiated hADSCs at 7 and 14 days after induction.ConclusionsIn conclusion, the findings suggest that CCh acts as an influential therapeutic factor in the field of neural regenerative medicine and research.
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Journal	Regenerative Therapy [Regen Ther]
Volume Number	23
DOI	10.1016/j.reth.2023.04.001
PubMed Central reference number	PMC10130343
PubMed reference number	37122359
e-ISSN	23523204
Language	English
Publisher	Japanese Society for Regenerative Medicine
Publisher Date	2023-04-15
Access Restriction	Open
Rights License	This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). © 2023 The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.
Subject Keyword	Adipose tissue-derived mesenchymal stem cells Calcium Carbachol Neural differentiation Tissue engineering
Content Type	Text
Resource Type	Article
Subject	Developmental Biology Biomaterials Biomedical Engineering