IF Staining of Human Primary Neurons

Primary Neurons are inputs or raw materials for cell based assays. When cells do not work as promised, there are multiple costs including lost time and potentially flawed data. Neuromics strives to provide easy to culture, potent and cost effective cells. Proving these capabilities is an ongoing activity for us. This includes testing these cells using our markers.

I wanted to share new immunofluorescence images. Here is a link to the protocol: staining primary neurons.

hN2 cells stained with our chicken polyclonal antibody to Vimentin, in red. Islands of Hn2 cells form after 4 days in culture forming beautiful flower like structures. Vimentin is a well established marker of early differentiating neuronal lineage cells. Taken with a 10X objective lens. Blue staining is the nuclear DNA.

hN2 cells grown in culture for 4 days and stained with our chicken polyclonal to MAP2, a marker of neurons. Differentiating cells show strong cytoplasmic staining for MAP2 . Blue stain is DAPI and reveals cell nuclei of some non neuronal cells in this culture.

hN2 cells grown in culture for 4 days and stained with our chicken polyclonal to Neurofilament light or low molecular weight chain NF-L, a marker of neurons. Many of the differentiating cells show strong cytoplasmic and clearly fibrillar staining for NF-L. Blue stain is DAPI and reveals cell nuclei of some non neuronal cells in this culture.

hNP1 Neural Progenitors Thawing+ Plating Protocol

This is a great video that shows thawing + plating protocol for our eSC Derived Human NP1 Neural Progenitors.

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.

STEMEZ(TM) hNP1 Human Neural Progenitors and hN2 Primary Neurons Differentiation and Expression

I have received a growing number of requests regarding differentiation and expression patterns of our STEMEZ^TM cells. This is intended help you gain a clearer understanding. The focus of this document is G-protein Coupled Receptor Expression Patterns. This information was presented by Dr. Steve Stice and his team and the 2008 Neuroscience Conference.


INTRODUCTION

Human embryonic stem cells and their progeny can provide a novel Distribution of detectable transcripts for three cell populations tissue source for understanding developmental pathways, pharmaceutical screening and tissue replacement therapies. G-protein coupled receptors(GPCRs) comprise the largest cell-surface receptor superfamily and are the largest class of drug targets. The study of GPCR signaling in hES cells allows signaling mechanisms to be studied in endogenously expressed receptors in non-transformed cells. We characterized GPCR transcript expression in three cellular populations at different developmental stages: WAO9 human embryonic stem cells, Wa09 derived STEMEZ hNP1 and differentiated hN2 cells maintained 1 week in culture.

Goal: To characterize GPCR transcript expression in human hES cell derived neural tissue

CONCLUSIONS
• hES cells displayed the widest array of GPCR transcripts, while neural progenitors displayed the most restricted population.
• The Frizzled (FZD) family of receptors were among the most abundantly expressed transcripts across all populations.
• Neural progentitors up-regulated GPCR transcripts important to brain angiogenesis, cell proliferation, neurogenesis and cell adhesion.
• Further differentiated hN2 cells displayed up-regulation of a wider population of transcripts including GPCRs involved with neurotransmission.
• Functional assays demonstrated responses to sphingosine-1-phosphate in both hNP1 and hN2 populations of cells.
• hES cells and their derived tissue provide a unique model to study endogenous GPCR signaling in non-transformed cells for drug screening applications and to further our understanding of GPCRs role in developmental pathways.

G-protein Coupled Receptor Expression Patterns Are Altered as Human Embryonic Stem Details(pdf - 367Kb). From Poster Presented at Neuroscience 2008 by Dr. Steve Stice et al.