Quality, High Titer Neuronal Markers

You Need Them; We got Them

We are recognized for both the number and quality of Neuronal Markers. Check out our many testimonials from satisfied customers.

Here's a sampling of recent publications.

Elisabet Garcia-Pino, Nikodemus Gessele and Ursula Koch. Enhanced Excitatory Connectivity and Disturbed Sound Processing in the Auditory Brainstem of Fragile X Mice. Journal of Neuroscience 3 July 2017, 2310-16; doi.org/10.1523/JNEUROSCI.2310-16.2017. ...chicken α-MAP2 (Neuromics; dilution 1:1000)...
Anna Lisa Gündner Claas Aiko Meyera, Stefan Aigner, Klaus Christensen, Christoph Patsch, Ravi Jagasia, Karlheinz Baumann, Marc Burcin. Generation of a homozygous GBA deletion human embryonic stem cell line. Stem Cell Research. Available online 11 July 2017. https://doi.org/10.1016/j.scr.2017.07.009 ...Neuronal differentiation marker, Ch-MAP2, 1:1000, Neuromics # CH22103 RRID:AB_2314763 ....Immunocytochemistry. Cells were cultured on PO/LAM coated 96-well imaging microplates (Falcon) and fixed with 4% paraformaldehyde (15 min, room temperature). Fixed cells were permeabilized with 0.2% Triton (Sigma) in phosphate-buffered saline (PBS) for 30 min and blocked for 1 h with PBS + 5% donkey serum (Merck Millipore). Primary antibodies (in PBS) were incubated overnight at 4 °C followed by three PBS washing steps. Secondary antibodies were incubated for 2 h at room temperature. Confocal images were acquired using a Leica TCS SP5 microscope (Leica Microsystems)....
Hayk Harutyunyan, Svetlana Sharoyan, Alvard Antonyan, Sona Mardanyan. Herb Preparations Improve the Viability of Hippocampal Cells Suppressed by Amyloid Beta (1-42) Peptide. World Journal of Pharmaceutical Sciences. 2017. ISSN (Online): 2321-3086......Antibodies for immunocytochemistry were purchased from Neuromics (USA)[....]Immunocytochemistry: Hippocampal cells were cultured in Poly-D-Lysine coated Nunc EasY Flasks. On the 4th day, the adhered cells were removed by trypsinisation (0.05% trypsin, 0.5 mM EDTA, pH 8.0). Cells were fixed on microscope slide by 3.7 % paraformaldehyde in PBS followed by methanol treatment. To block a nonspecific antibody binding, samples were pre-treated with goat serum (Sigma) for 30 minutes. To determine the types of cells, constituting the obtained cell culture, the different aliquots of the culture were incubated for 60 min at room temperature with the primary antibodies against rabbit anti-rat Neurofilament (NF), chicken anti-rat GFAP and Nestin...

Images: Cells grown from adult rat brainLarge cell in middle is stained with mouse monoclonal to NF-L clone DA2 (green). Another type of neuronal lineage cell was stained with rabbit polyclonal to alpha-internexin (red). These cells were mitotic but had several characteristics of neurons. Rat spinal cord homogenate showing the major intermediate filament proteins of the nervous system (lane 1). The remaining lanes show blots of this material stainted with various antibodies including: NF-H, NF-M,, NF-L,, NF66 and GFAP 

We will continue to post the latest and greatest news.

Stem Cell "Stemness" and Differentiation States

Markers Play a Key Role

Our Stem Cell Markers are widely used and frequently published. These play an important role for understanding the molecular events that take place during the differentiation of human pluripotent cells. They are essential for the protocols aimed at insuring the generation high of quality differentiated cells.

Here's an excellent study on the determining the state of differentiating stem cells: Julio Castaño, Cristina Morera, Borja Sesé, Stephanie Boue, Carles Bonet-Costa, Merce Martí, Alicia Roque, Albert Jordan, Maria J. Barrero. SETD7 Regulates the Differentiation of Human Embryonic Stem Cells. Published: February 18, 2016DOI: 10.1371/journal.pone.0149502...SOX2 (Neuromics GT15098)...

Figure: SETD7 is expressed at very low levels in pluripotent human cells and induced during differentiation. (A) Average rank of the top 700 most differentially expressed genes between pluripotent (iPSCs or ESCs) and fibroblasts, including those upregulated in pluripotent cells (left panel) and upregulated in fibroblasts (right panel). (B) SETD7 mRNA levels in human ESCs grown under self-renewal conditions (UndES[4]), in vitro differentiated human ESCs (DifES[4]), human fibroblasts (HFF), two lines of human keratinocytes (HEK1 and HEK2) and two lines of iPSCs generated from keratinocytes ([H]KiPS4F and KiPS4F1). Mean and standard deviation of three technical replicates is shown. Induction of SETD7 mRNA levels during ES[4] differentiation was confirmed in more than four independent differentiation experiments. (C) Western blot showing SETD7 protein levels in pluripotent and somatic cells. Loading control beta actin (ACTB) is also shown. (D) Western blot showing protein levels of SETD7, AFP, OCT4 and SOX2 in under self-renewing conditions and in vitro differentiated human ESCs. Loading control alpha tubulin (TUBA) is also shown. One representative experiment out of three is shown. (E) Genomic visualization of the levels of H3K72me3, H3K4me3, H3K4me2, H3K36me3 and RNA polymerase II (Pol II) in the human embryonic stem cell line H1 around the SETD7 gene according to ENCODE. A non-methylated CpG island is depicted in green. (F) Levels of H3K4me2 and H3K27me3 at SETD7 gene promoter region (27 bp upstream of the transcription start site) in pluripotent and somatic cells determined by chromatin immunoprecipitation (ChIP) and ploted relative to the input. IgGs wer used as negative control. Bars show the mean and standard deviation of three independent immunoprecipitations...doi:10.1371/journal.pone.0149502.g001

Neuromics is pleased to provide many different stem cell related options for your cell based assays. I will continue to post updates.

Everything You Wanted to Know About Stem Cells

Unforgettable way to learn about Stem Cells.

Rap on Dr. Jonathan Garlick Rap on.

If you are doing stem cell related research, check out our:

hNP1™ Human Neural Progenitor & hN2™ Neuron Discovery Kits Derived from H9 (WA09) ECSs-Consistent, Easy to Use & Cost Effective	Neural Stem Cells Media
hMPro™ Human Mesenchymal Progenitors (hMPCs) hESC Derived	Human Mesenchymal Stem Cells (hMSCs) hMSCs Derived from Umbilical Cord Blood
MSCGro™ Mesenchymal Stem Cell Media Proven and Potent Culture Growth and Differentiation Media for Mesenchymal Stem Cells (MSCs)	Cell Cryopreservation Media Maximizes cell recovery, attachment and growth
Expansion/Differentiation Kits SC to Oligodendrocytes and Dopaminergic Neurons Kits	Bioluminomics™ In Vitro Assays Brought to you by HemoGenix®
3-D Cell Based Assay Solutions Nanofibers, Hydrogels and Extracellular Matrix (ECM) Proteins	Stem Cell Research Antibodies
Stem Cell Research Proteins	Rat Neurosphere Neuroprogenitor Tissue Tissue provided live, unseparated, fresh from E18 rat cortex/hippocampus including subventricular zone.

eSC Derived human Mesenchymal Progenitor Pubs

We have added hMPro^TM Mesenchymal Progenitors to our Stem Cell Research Reagents category. These progenitors coupled with our UBC derived Human Mesenchymal Stem Cells give more options to basic and drug discovery researchers. Since they serve as potent research tools, it is a strategy of ours to continue to broaden our offerings in this products category.

Publications matter when assessing the capabilities of bio-reagents. I would like to share several that prove the utility of these cells:  David L. Simpson, Nolan L. Boyd, Sunjay Kaushal, Steve L. Stice, Samuel C. Dudley Jr. Use of human embryonic stem cell derived-mesenchymal cells for cardiac repair. Biotechnology and Bioengineering Volume 109, Issue 1, pages 274–283, January 2012. DOI: 10.1002/bit.23301

Nolan L. Boyd, Ph.D., Kelly R. Robbins, Ph.D., Sujoy K. Dhara, D.V.M., Ph.D., Franklin D. West, Ph.D., and Steven L. Stice, Ph.D. Human Embryonic Stem Cell–Derived Mesoderm-like Epithelium Transitions to Mesenchymal Progenitor Cells. Tissue Eng Part A. 2009 August; 15(8): 1897–1907. Published online 2009 January 15. doi: 10.1089/ten.tea.2008.0351.

Images: (A) Phase contrast image of hMPro™ mesenchymal progenitor cells in culture. (B) Confocal image of hMPro™ cells stained for the early smooth muscle marker (αSMA;green), F-actin (red) and nuclei (blue). Exposure to 10ng/mL of transforming growth factor beta 1 (TGF-β1) for 12 days induces expression of alpha smooth muscle actin (αSMA) in WA09-derived mesenchymal progenitor cells, suggesting their ability to differentiate along the smooth muscle lineage.

These cells have the ability to:

•Form adherent monolayers – ideal for 96- and 384-well formats for high throughput and high content cell-based assays.
•Differentiate into a wide variety of mesenchymal subtypes, including osteogenic and chondrogenic lineages (but not adipogenic)– ideal for drug discovery for a wide variety of biological targets and basic research in bone, cartilage, metabolic and immunological diseases.
•Passage up to 10X.

I will keep you posted on new applications.

100% Animal Free FGF Basic

New ISO-kine^TM FGF basic-only 39 USD-10 ug.

Fibroblast Growth Factor (FGF) Recombinant Proteins are widely used as tools in Stem Cell Research. They have the capabilities to catalyze growth and differentation as well as maintain cell stasis. We continue to offer new FGFs to make sure we meet the unique demands of our cutsomers and collaborators  .

Our E. Coli derived Human FGF (146 aa) recombinant protein is our #1 seller. As Stem Cell research moves from the bench top to the bedside, there will be a growing need for 100% animal free proteins. We are please to announce we have added a 100% animal free FGF (146 aa) basic that has bio-activity comparable to our top selling options

Image: ISOKine^TM bFGF vs our potent and proven e-coli derived bFGF.

I anticipate offering more ISOKine^TM Bio-risk free Stem Cell Research growth factors to meet growing demand. This growth will be driven by their unique advantages. These include low protease activity and secondary metabolite content and simple protein content all of which aid in downstream processing. These products also have the G.R.A.S. (Generally Recognised As Safe) status from FDA.

Stem Cell Differentiation Flow Charts

Installment #1: Mesoderm Derived Stem Cells

We would like to thank our partners and customers for their input on how to best add value to our Stem Cell Research Reagents. Based on this input, we are in the process of building a new website focused on providing stem cell technical content. Better content should help catalyze better research and drug discovery related decisions.

Here're several stem cell differentiation flow charts that map the genesis of pluripotent and multipotent cells and identify the cell types that result from their differentiation.

This chart was generated using input and publications generously provided by Dr. Henry Young, Tenured Full Professor at Mercer University School of Medicine.

I anticipate the launch date for our Stem Cell Related website to occur in early Q3 2012.

In the meantime, we will posting relevant data, publications and other information here. This will include protocols and related growth factors for generating specific cell types like dopaminergic neurons from neural progenitors. Stay tuned.

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.

Lineage Selection-Neural Stem Cells for SC Grafts

Our Neural Progenitor Markers keep moving up in the "hit parade". These markers are important for lineage selection. This selection is essential to circumvent the possibility of tumor formation and facilitate the safe translation of ES-based therapies to humans.

Here's a recent pub referencing use of several of our markers for selecting or confirming lineage: J. Simon Lunn, Crystal Pacut, Emily Stern, Stacey A. Sakowski, J. Matthew Velkey, Sue O'Shea, Eva Feldman. Intraspinal transplantation of neurogenin-expressing stem cells generates spinal cord neural progenitors. dx.doi.org/10.1016/j.nbd.2011.12.044...Tuj1 (Neuromics, 1:1000), Nestin (Neuromics, 1:500)...
Highlights: Expression of appropriate transcription factors is one approach to direct the differentiation of ES cells towards a specific lineage and stop proliferation. Neural differentiation can be initiated in ES cells by expression of Neurogenin1 (Ngn1). In this study we investigate the effects of controlled Ngn1 expression on mouse ES (mES) cell differentiation in vitro and following grafting into the rat spinal cord. In vitro, Ngn1 expression in mES cells leads to rapid and specific neural differentiation, and a concurrent decrease in proliferation. Similarly transplantation of Ngn1-expressing mES cells into the spinal cord lead to in situ differentiation and spinal precursor formation. These data demonstrate that Ngn1 expression in mES cells is sufficient promote neural differentiation and inhibit proliferation, thus establishing an approach to safely graft ES cells into the spinal cord.

Image: Neural progenitors were labeled with anti-rat Nestin polyclonal antibody (Cat#:GT15114) and stained with conjugated donkey anti-goat secondary anti-body (green). Differentiated neurons were labeled with neuron-specific mouse anti-β-III tubulin/ Tuj1-(Cat#MO15013 monoclonal antibody (red). Nuclei were stained with DAPI (blue).

I will continue to post updates on the application of Neuromics' Stem Cell Markers

Mouse epiSCs Into Myelinating Cells

This study published recently in Nature Methods hit my radar scope becaused it referenced use of our widely used and frequently published stem cell marker Tuj 1 (Neuron-specific class III beta-tubulin): Fadi J Najm, Anita Zaremba, Andrew V Caprariello, Shreya Nayak, Eric C Freundt, Peter C Scacheri, Robert H Miller & Paul J Tesar. Rapid and robust generation of functional oligodendrocyte progenitor cells from epiblast stem cells. Nature Methods (2011) doi:10.1038/nmeth.1712.

Dr. Paul Tesar and his team at Case Western University demonstrated the ability to convert pluripotent epiblast stem cells into pure populations of myelinating cells, called oligodendrocyte progenitor cells (OPCs). First, stem cells in a petri dish are treated with molecules to direct them to become the most primitive cells in the nervous system. To produce OPCs, these primitive cells are treated with a defined set of proteins. The cells were cultured on laminin and treated withh apporopriate growth factors. The OPCs were nearly homogenous and could be multiplied to obtain more than a trillion cells.

The OPCs were treated with thyroid hormone, which is key to regulating the transition of the OPCs to oligodendrocytes. The result was the OPCs stopped proliferating and turned into oligodendrocytes within four days.

These methods could used to potentially produce stable and pure populations of human OPCs in a significant enough number to treat patients with demyelinating diseases such as multiple sclerosis and cerebral palsy.

SOX2 and Initiation of Breast Tumors

I consider it a feather in Neuromics' cap when our reagent(s) are referenced in a Nature Journal. More importantly it enables me to keep the pulse on novel and important discovery.

This pub references use of one of our SOX2 Antibody and comes from Dr. Angel García Martín and his Team at INBIOMED: O Leis, A Eguiara, E Lopez-Arribillaga, M J Alberdi, S Hernandez-Garcia, K Elorriaga, A Pandiella, R Rezola and A G Martin. Sox2 expression in breast tumours and activation in breast cancer stem cells. Oncogene , (8 August 2011) | doi:10.1038/onc.2011.338.

The important insight from this study is: "Over-expression of Sox2 increased mammosphere formation, effect dependent on continuous Sox2 expression; furthermore, Sox2 knockdown prevented mammosphere formation and delayed tumour formation in xenograft tumour initiation models. Induction of Sox2 expression was achieved through activation of the distal enhancer of Sox2 promoter upon sphere formation, the same element that controls Sox2 transcription in pluripotent stem cells. These findings suggest that reactivation of Sox2 represents an early step in breast tumour initiation, explaining tumour heterogeneity by placing the tumour-initiating event in any cell along the axis of mammary differentiation."

Could these findings ultimately lead to a better diagnostic for Breast Cancer? I'll keep you posted.

Lab Highlights: Breast cancer is the most frequent cancer in women making up to 20% of all tumours diagnosed in women, with 1 million new cases diagnosed every year worldwide (16,000 new cases only in Spain). Worlwide it causes over 350,000 deaths with an increasing tendency. Breast cancer stem cells show the phenotype CD44+/CD24low/-Lin- though only a fraction of this population has the capacity to initiate tumours. Therefore a complete and precise description of the breast cancer stem cells is lacking.

The focus of this laboratory is to identify, isolate and culture breast cancer stem cells from natural breast tumours and compare at the molecular level with normal mammary stem cells. This research involves the stablishment of both in vitro and in vivo functional assays and the molecular characterization (both genomic and proteomic) of breast cancer stem cells to define the mechanisms responsible for its transformed phenotype.

High Content and High Throughput Toxicity Screening

Kits designed for Drug Discovery and Development

Our customers have been impressed with the capablities of our in vivo and in vitro apoptosis and toxicity kits. Here's a recent pub referencing use of one of our FLICA™ in vitro Caspase Kits: Giovanna Grandinetti, Nilesh P. Ingle, and Theresa M. Reineke. Interaction of Poly(ethylenimine)–DNA Polyplexes with Mitochondria: Implications for a Mechanism of Cytotoxicity. Mol. Pharmaceutics, Article ASAP Publication Date (Web): June 23, 2011 Copyright © 2011 American Chemical Society...inhibitor of caspases (FLICA) specific for caspase-9 (Neuromics, Inc)...
Images: Jurkat cells were treated with 1 µM staurosporine for 3 hours to induce caspase 9 activity (top), or were treated with a control (bottom). Both populations were incubated with ICT’s green FAM-LEHD-FMK FLICA™ caspase 9 reagent. DIC images were taken of both samples. Almost all cells in the induced sample (top) fluoresce green therefore they have activated caspase 9. None of the control cells (bottom) fluoresce green, therefore they do not have activated caspase 9 Courtesy of Dr. Brian Lee, ICT

Neuromics is pleased to announce the addition of HemoGenix® Predictive in vitro Toxicity and Apoptosis Kits:

Solutions for predictive in vitro toxicity and apoptosis are now important than ever. These kits enable you to do high throughput and high content screening of stem cells, progenitors and primary cells.
Kit options include:

•LumiSTEM™-96 iPS and LumiSTEM™-iPS HT Assays to Study Induced Pluripotent Stem Cells (iPS) and Toxicity to iPS Cells and Cells Derived from iPS Cells.

•LUMENESC™-Tox HT (LUMENESC™-96 Tox and LUMENESC™-384 HT). A Toxicity Screening and Testing Platform for Cells of the Mesenchymal Stem/Stromal Cell System.

•HALO®-Tox HT Predictive Hemotoxicity Platform using HALO®-96 Tox and HALO®-384 HT. A Highly Predictive, In Vitro Stem and Progenitor Cell Hemotoxicity Screening and Testing Platform for all Stages of Drug Development and Xenobiotic.

I will continue to post regarding progress.

Potent and Cost Effective Cell Based Assays

I have had many conversations with basic and drug discovery researchers on improving cell based assays. Here's the wish list:

• More potent cells/media
• More accurate analytic tools-quatititative and reproducible results
• Ability to use cells and tools in high throughput/high content screening.
• Cost effectiveness

This wish list is front and center in determining the cells/media and related tools we add to Neuromics' offerings. We are pleased to announce the addition of our Hemogenix's Bioluminomics™ In-Vitro Cell Assays, MSCGro™ Mesenchymal Stem Cell Media and Umbilical Cord Blood derived hMesenchymal Stem Cells.

These provide quantitation, not subjectivity. It includes assay calibration and standardization. It means assay validation. It produces results you can trust and rely on. It means innovation and flexibility. It is advanced technology that is fast to learn, easy to use and above all, cost effective.

Assays options:

• LumiSTEM™ for Primary Stem and Explanted Cells, Stem and Other Cell Lines-Selecting LumiSTEM™ Assays(pdf - 418Kb)
• LUMENESC™ for Mesenchymal Stem/Stromal Cell System-Selecting LUMENESC™ Assays(pdf - 984Kb)
• HALO® for for Human Lympho-Hematopoietic Stem and Progenitor Cell Research.-Selecting HALO™ Assays(pdf - 1.7M)
• Biolumonics Basic Procedures(pdf - 323Kb) 

Available Cells:

Primary Neurons and Astrocytes-Primary human, rat and mouse neurons and astrocytes
STEMEZ^TM Human Neural Progenitor Neuron Discovery Kits-Derived from H9 (WA09) ECSs-Consistent, Easy to Use & Cost Effective
Human Mesenchymal Stem Cells (hMSCs-hMSCs derived from pancreas and umbilical cord blood
Mammalian Cell Lines

Media:
STEMEZ(TM) hN2 Human Neurons Culture Media
MSCGro™ Mesenchymal Stem Cell Media
NbActiv4

I will continue to post customer input and related data on Neuromics' Cell Based Assay Tools.

H9-WA09 Derived Human Neural Progenitors and Neurons Pricing

Neuromics offers stable, potent and well characterized STEMEZ (TM) Human Neural Progenitor & Neuron Discovery Kits. These are derived from NIH registered human ES cell line H9 (WA09).

Details on the capabilities of these cells are detailed in a variety of publications. They work! As a result, I am trying to make it easier to justify using them when and where they are needed. In addition to having frequent updates on new references, methods and data, I want to offer the best pricing possible.

In researching pricing of cells derived from the same parental line and having similar characteristics, I noticed prices ranging from 995 to 2800 USD. Our pricing starts at 695 USD and we offer deeper discounting if required. I do not want price to be a barrier. Here's some sample data on these cells.

Images: Neural phenotypes derived from hN2 cell lines. (A) Phase contrast image of differentiated culture. (B) Network including post-mitotic motoneurons (HB9). (C) Cholinergic neuron. (D) Tuj-1 positive cells that are DAT-positive (dopamine transporter; closed arrow) and DAT-negative (open arrow). (E) Gabaergic neurons, inset illustrates GABA in axon, but not the dendrites (arrow).

G-protein Coupled Receptor Expression Patterns Are Altered as Human Embryonic Stem Details-From Poster Presented at Neuroscience 2008 by Dr. Steve Stice et al.
hN2 Cells-Electro Phys Data.

Related Products:

• All Stem Cell Research Reagents
• Primary Neurons and Astrocytes

I wish exciting and rewarding discoveries.

Stems Cells and SCI

This is fascinating and encouraging to sufferers of Spinal Cord Injury:

ScienceDaily (Oct. 9, 2010) — Researchers at Karolinska Institutet have shown how stem cells, together with other cells, repair damaged tissue in the mouse spinal cord. The results are of potential significance to the development of therapies for spinal cord injury: http://www.sciencedaily.com/releases/2010/10/101008082736.htm#

Isolation of Medulloblastoma Stem Cells (Video Protocol)

I am pleased to present this excellent video. It also has excellent images using key stem cell markers as the cells undergo differentiation:

JoVE: Isolation, Enrichment, and Maintenance of Medulloblastoma ... Sep 1, 2010 ... GFAP antibody, Neuromics, CH22102, Chicken, 1:1000. Tuj1 antibody, Sigma, T5076, Mouse, 1:2000. NeuN antibody, Millipore, MAB377, Mouse, ... www.jove.com

Otx2 (Orthodenticle Homeobox 2) and Parkinson's Disease

Our Otx2 Antibody is a potent marker for Human, Mouse and Rat Midbrain Dopamanergic Progenitors.

This is confirmed by a recent publication by Dr. Ole Isaacson et al:

Chee Yeun Chung, Pawel Licznerski, Kambiz N. Alavian, Antonio Simeone, Zhicheng Lin, Eden Martin, Jeffery Vance and Ole Isacson. The transcription factor orthodenticle homeobox 2 influences axonal projections and vulnerability of midbrain dopaminergic neurons. Brain Advance Access published online on June 23, 2010 Brain, doi:10.1093/brain/awq142... anti-Otx2 (Neuromics, 1:500)...

Abstract: Two adjacent groups of midbrain dopaminergic neurons, A9 (substantia nigra pars compacta) and A10 (ventral tegmental area), have distinct projections and exhibit differential vulnerability in Parkinson’s disease. Little is known about transcription factors that influence midbrain dopaminergic subgroup phenotypes or their potential role in disease. Here, we demonstrate elevated expression of the transcription factor orthodenticle homeobox 2 in A10 dopaminergic neurons of embryonic and adult mouse, primate and human midbrain. Overexpression of orthodenticle homeobox 2 using lentivirus increased levels of known A10 elevated genes, including neuropilin 1, neuropilin 2, slit2 and adenylyl cyclase-activating peptide in both MN9D cells and ventral mesencephalic cultures, whereas knockdown of endogenous orthodenticle homeobox 2 levels via short hairpin RNA reduced expression of these genes in ventral mesencephalic cultures. Lack of orthodenticle homeobox 2 in the ventral mesencephalon of orthodenticle homeobox 2 conditional knockout mice caused a reduction of midbrain dopaminergic neurons and selective loss of A10 dopaminergic projections. Orthodenticle homeobox 2 overexpression protected dopaminergic neurons in ventral mesencephalic cultures from Parkinson’s disease-relevant toxin, 1-methyl-4-phenylpyridinium, whereas downregulation of orthodenticle homeobox 2 using short hairpin RNA increased their susceptibility. These results show that orthodenticle homeobox 2 is important for establishing subgroup phenotypes of post-mitotic midbrain dopaminergic neurons and may alter neuronal vulnerability.

Image: Characterization of the human neuroectodermal precursors. Otx2 Staining of forebrain-midbrain rosettes (dilution 1:1000).

All Publications Referencing Neuromics Purified Goat Polyclonal Otx2

Related Reagents to Consider:

• Otx2-Mouse 
• Stem Cell Research Reagents

Dr. Steve Stice to Present the Power of StemEZ Neural Cells

Dr. Steve Stice to Present the Power of StemEZ Neural Cells

Posted using ShareThis

I have profiled Steve Stice's research here. The focus has been the excellent research results he and his team at ArunA Biomedical have generated with STEMEZ(TM) hN2 Human Neurons and hNP1 Human Neural Progenitors.

The story continues. He will be presenting the latest at the 9th Annual World Pharmaceutical Congress in Philadelphia, June 14. Topics include: using these neural cell lines to study neurotoxicity in cell-based assays and disease modeling. Recent work conducted in outside laboratories demonstrates that these lines are more sensitive to environmental toxicants than traditional cellular models.

Sample high throughput assay applications:

• Cell morphology and neurite outgrowth


• Cell signaling and transcription factor expression


• Receptor and ion channel function


• Cytotoxicity
  
• Apoptosis, genotoxicity and DNA damage
  

These capabilities has been confirmed by our customers. I look for the use of the STEMEZ cell lines to continue to grow as researchers discover their value in Drug Discovery and Basic Neuroscience capabilities.

STEMEZ hNP1 Neural Progenitors Now Available

We are pleased to announce the introduction of our STEMEZ ^TMhNP1 Human Neural Progenitors Discovery Kit. The kit was developed by our partner, ArunA Biomedical.

Description/Data:
STEMEZ^TM hNP1 Human Progenitors are fully differentiated are derived as adherent cells from hESC WA09 line. The cells are shipped frozen in a vial with 1 x 10⁶ cells; once thawed they should be immediately plated on a Matrigel (or other suitable extracellular matrix protein)-coated dish and maintained in the accompanying serum-free medium. The neurons should be used within 14 days of thawing. These cells have the capabilities to:

• Display immunoreactive properties consistent with neural precursors and can be maintained in a proliferative state in monolayer cultures.
• Differentiate under serum-free conditions into neuronal phenotypes with functionally responsive transmitter receptors in vitro .
• Maintain multiple neural phenotypes in long term serum-free culture.
• Can be directed to alter phenotypic characteristics under different media conditions.

Applications: Gene Expression Analysis Western blotting, Flow Cytometry, Immunocytochemistry, FACS sorting, DNA Microarray, RT-PCR, Neurite Outgrowth assays, FLIPR calcium assays, cytotoxicity, and second messenger signaling.

Images: STEMEZ^TM hNP1 Human Neural Progenitor Cells are grown as monolayers (A), are karyotypically normal (B) and express NSC markers, Nestin Mouse and Sox-2 (C, D, E). Nuclei of the cells were visualized with DAPI (blue). The Sox-2 transcription factor is co-localized with the DAPI (blue) staining in the nucleus (F).

STEMEZ hNP1 FAQs

Note: STEMEZTM NP1 Progenitors and hN2 Human Neurons can be genetically manipulated with GFP reporters without altering the function and differentiation potential of these cells. In contrast to many stem cell sources, the reporter genes are not readily silence in the neural cultures. See: doi:10.1089/ten.tea.2009.0155.

DNA fingerprint cells: The loci match the DNA fingerprint pattern for the H9 (NIH designation, WA09) HESC line as published in http://stemcells.nih.gov/research/nihresearch/scunit/.

Viral tests cells: This lot was derived from the H9 hESC line that has been tested for Hepatitis B, Hepatitis C, HIV-1, HIV-2, HTLV-I/II, HSV1, HSV2, EBV, and CMV. The H9 cell line has been tested and shown to be negative. (Tests performed by GIVF Laboratories).

Related Reagents:

• STEMEZ(TM) hN2 Human Neurons Discovery Kit


• Neuron/Glial Marker Antibodies


• Neurotrophins and Growth Factor Antibodies


• Fibroblast Growth Factor Proteins


• Neurotrophins-Neuron/Glial Marker Proteins


• Stem Cell Reagents


• Transfection Reagents

Potent Stem Cell Markers

The potency of our Stem Cell Research Reagents is confirmed by the growng list of publication referencing them.

Here're the latest:

Saishu Yoshida, Takako Kato, Takao Susa, Li-yi Cai, Michie Nakayama, Yukio Kato. PROP1 coexists with SOX2 and induces PIT1-commitment cells. Biochemical and Biophysical Research Communications, Volume 385, Issue 1, 17 July 2009, Pages 11-15
... SOX2 (1:500 dilution, Neuromics, Edina, MN, USA)...

Saravanan Karumbayaram, Bennett G. Novitchb, Michaela Patterson, Joy A. Umbach, Laura Richter, Anne Lindgren, Anne E. Conway, Amander T. Clark, Steve A. Goldman, Kathrin Plath, Martina Wiedau-pazos, Harley I. Kornblum, William E. Lowry. Directed Differentiation of Human-Induced Pluripotent Stem Cells Generates Active Motor Neurons. Stem Cells Vol. 27 No. 4 April 2009, pp. 806 -811. doi:10.1002.
... mouse anti-Nestin (Neuromics)...

Patrizia Rubini, Javorina Milosevic, Johannes Engelhardt, Mahmoud Al-Khrasani, Heike Franke, Attilla Heinrich, Beata Sperlagh, Sigrid C. Schwarz, Johannes Schwarz, Wolfgang Nörenberg and Peter Illes. Increase of intracellular Ca2+ by adenine and uracil nucleotides in human midbrain-derived neuronal progenitor cells. Cell Calcium. Volume 45, Issue 5, May 2009, Pages 485-498.
...rabbit anti-P2Y2 (1:1000; Neuromics, Edina, MN, USA...