Hello

I prefer methods of Woodbury and colleagues . Where Subconfluent cultures of rat and human MSCs were maintained in DMEM/20% FBS. Twenty-four hours prior to neurona linduction, media were replaced with preinduction media consisting of DMEM/20% FBS/1 mM b-mercaptoethanol (BME). To initiate neuronal differentiation, the preinduction media were removed, and the cells were washed with PBS and transferred to neuronal induction media composed of DMEM/1–10 mM BME.

In later experiments DMEM/2% dimethylsulfoxide (DMSO)/200 mM butylated hydroxyanisole (BHA) was utilized as the neuronal induction media. Cells were fixed for immunocytochemistry at times ranging from 30 min to 6 days postinduction.

in attachment the full artical , Good Luck

This is controversial and the resulting 'neural' cells, if neural at all, are probably not functional. There is lots of literature on this, most of which is of poor quality so be careful to properly assess the presented data. The efficiency of current protocols are low and associated with high levels of cell death. Be sure to notice the frequency of positive cells in the images shown to you, and bear in mind that these images will bias towards some of the 'better' regions, as well as the problem that surface marker expression is not conclusive of differentiation into the associated cell type.

The woodbury protocols Saleh Alkarim suggest were the first to report this 'transdifferentiation', I think, along with Sanchez-Ramos et. al. 2000. But be aware that a later publication criticised the findings (attached).

Article Induction of bone marrow stromal cells to neurons: Different...

I agree with Nathan Sweeney. We used 4 years ago the protocol with DMSO etc..And yes, MSCs looked like "neurons", expressed low level of nNOS, but died after 24 hours. Later I found the paper of Neuhuber et al."Reevaluation of In Vitro Differentiation

Protocols for Bone Marrow Stromal Cells: Disruption of Actin Cytoskeleton Induces

Rapid Morphological Changes and Mimics Neuronal Phenotype" where they demonstrated that fibroblasts look also like neurons if the differentiation protocoll with DMSO is used: http://neurobio.mcphu.edu/FischerWeb/PDF/JNeurosciResearch.2004.77.192.pdf

That is a tricky question, because mesodermal stem cells are not in the same developmental lineage as neural tissues [Adult stem cells. Anat. Rec. 276A:75-102, 2004]. Since most studies I am aware of did not use 100% pure clonal populations of MSCs, but rather mixed populations of primary isolates they actually did not know the identity of their starting population of cells [Neuronal differentiation of stem cells isolated from adult muscle. J Neurosci Res 69:894-907, 2002; Adult reserve stem cells and their potential for tissue engineering. Cell Biochem Biophys, 40(1):1-80, 2004]. I suspect that in their MSC population were also neural progenitor cells, ectodermal stem cells, pluripotent stem cells and/or totipotent stem cells [Pluripotent Stem Cells, Endogenous versus Reprogrammed, a Review. MOJ Orthop Rheumatol 1(4): 00019, 2014. https://lnkd.in/eUYkgU6" MOJ ISSN: 2374-6939MOJOR]. One needs to know the composition and identity of the cells they are working with before they can come to any reasonable conclusions concerning their studies.