Fibroblasts into Neurons

Via Activation of Oct4

We have a variety of stem cell, progenitor and neuron markers that are often referenced in publication. There represent a qualitative way to determine the state of stem cells as they differentiate.

Here're researchers take Fibroblasts and differentiate them into various progenitors. They were able to further differentiate these progenitors into astrocytes and neurons by: For neuronal differentiation, after 2 days of incubation with 20 nM reversine, cells were then treated with 0.5 μM all-trans-retinoic acid (RA) in serum-free DMEM/F-12 medium supplemented with the ITS for 2 days and switched into serum-free medium in the absence of RA with replacement of the medium every 2-3 days. 

Our NSE and GFAP were used to confirm this differentiation.

To learn more see: Xiangchen Li, Yu Guo, Yaxin Yao, Jinlian Hua, Yuehui Ma, Changqing Liu, Weijun Guan.Reversine Increases the Plasticity of Long-Term Cryopreserved Fibroblasts to Multipotent Progenitor Cells through Activation of Oct4. International Journal of Biological Sciences 2016; 12(1): 53-62. doi: 10.7150/ijbs.12199

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Should you ever have questions or issues, I am at your "beck and call". Do not hesitate to contact me directly. Pete Shuster, CEO and Owner, Neuromics, 612-801-1007 or pshuster@neuromics.com. Thank you!

Neural Progenitors and Cool Science

Titania Nanotubes and Neural Prostheses

I am always on the hunt for the use of our Neural Progenitor Markers in cool and important human health applications. Here our Nestin Antibody is use to access the growth and maintenance of C17.2 neural stem cell line cultured in these nanotubes: Jonathan A. Sorkin, Stephen Hughes, Paulo Soares, Ketul C. Popat. Titania nanotube arrays as interfaces for neural prostheses. Materials Science and Engineering: C Volume 49, 1 April 2015, Pages 735–745. doi:10.1016/j.msec.2015.01.077

Image: Nestin staining eSC Derived hNP1 Human Neural Progenitors. Cells were stained using goat anti-mouse Alexa Fluor 488 secondary antibody (green) (Molecular Probe, A-11001) and counterstained with DAPI (blue).

Abstract: Neural prostheses have become ever more acceptable treatments for many different types of neurological damage and disease. Here we investigate the use of two different morphologies of titania nanotube arrays as interfaces to advance the longevity and effectiveness of these prostheses. The nanotube arrays were characterized for their nanotopography, crystallinity, conductivity, wettability, surface mechanical properties and adsorption of key proteins: fibrinogen, albumin and laminin. The loosely packed nanotube arrays fabricated using a diethylene glycol based electrolyte, contained a higher presence of the anatase crystal phase and were subsequently more conductive. These arrays yielded surfaces with higher wettability and lower modulus than the densely packed nanotube arrays fabricated using water based electrolyte. Further the adhesion, proliferation and differentiation of the C17.2 neural stem cell line was investigated on the nanotube arrays. The proliferation ratio of the cells as well as the level of neuronal differentiation was seen to increase on the loosely packed arrays. The results indicate that loosely packed nanotube arrays similar to the ones produced here with a DEG based electrolyte, may provide a favorable template for growth and maintenance of C17.2 neural stem cell line.

I will continue to how our stem cell solutions are used in cool, new discoveries.

Ready for Prime Time Stem Cell Markers

Researchers frequently reference use of Neuromics' Stem Cell Markers in publications. This is an important affirmation for us as these tools are critical for determining the differentiation state of Stem Cells. In this publication, the authors use our Mouse Monoclonal Nestin Antibody to understand the mechanisms underlying neural progenitor differentiation and neuronal fate. This understanding is an important precursor for using these cells in Regenerative Medicine: Serafí Cambray, Charles Arber, Graham Little, Antonios G. Dougalis, Vincenzo de Paola, Mark A. Ungless, Meng Li and Tristan A. Rodríguez. Activin induces cortical interneuron identity and differentiation in embryonic stem cell-derived telencephalic neural precursors. Nature Communications 3, Article number: 841 doi:10.1038/ncomms1817. Received 10 January 2011 Accepted 29 March 2012 Published 15 May 2012.

In this study, the authors show that Activin provides telencephalic neural precursors with positional cues that specifically promote the acquisition of a calretinin interneuron fate by controlling the expression of genes that regulate cortical interneuron identity. This work demonstrates a novel means for regulating neuronal differentiation and specification of subtype identity.

Images: (a)immunostaining (left panels) and quantifications (right panel) indicating that Shh promotes and cyclopamine inhibits proliferation in neural precursors (Nestin+/β-III-tubulin+ cells in cyclopamine 49±4.3/35.1±1.8%, Shh 80.3±3.2/14.8±0.7% and control cultures 68.4±7.2/20.1±2.8%; n=3, mean±s.e.m.). (b) Relative expression levels of Gli1 and Ptch1 during the first 5 days of Activin or control treatment. (c) Immunoblot analysis of Gli1 and Cyclin D1 levels during the first 4 days of Activin or control treatment illustrating how Activin represses the expression of these proteins. (d) Normalized mRNA levels of Gli1 and Ptch1 after 24 h exposure to the indicated conditions illustrating how Shh induces the expression of these genes and Activin inhibits their expression (n=3, mean±s.e.m. Student's t-test. *P<0.005 and **P<0.05). ESCs were differentiated for 5 days as a monolayer, then replated into poly-D-lysine/laminin-coated dishes and cultured in NBB27 media (controls), NBB27+10 ng ml−1 Activin, NBB27+100 ng ml−1 Shh, NBB27+10 μm cyclopamine or NBB27 + 10 ng ml−1 Activin + 100 ng ml−1 Shh. Scale bar=50 μm.

Image: 

Image: Model for how Activin induces the differentiation and CGE fate in telencephalic neuronal precursors.


The protocol described in this manuscript represents a method to obtain an enriched source of calretinin interneurons from both mouse and human ESCs. Therefore, our work significantly contributes to the aim of generating the diverse neuronal subtypes required for the safe and successful use of ESCs in regenerative medicine.

The capabilities of stem cells markers matter in developing protocols for use in Regenerative Medicine.