Tracking STEMEZ hNP1 Progenitor Cell Fate

We would like to promote an important new discovery using our STEMEZ(TM) hNP1 Human Neural Progenitors Discovery Kit.

Human embryonic stem cell–derived neural progenitors (NP) present an important tool for understanding human development and disease. Optimal utilization of NP cells, however, requires an enhanced ability to monitor these cells in vitro and in vivo. Here we report production of the first genetically modified self-renewing human embryonic stem cell–derived NP cells that express fluorescent proteins under constitutive as well as lineage-specific promoters, enabling tracking and monitoring of cell fate. Nucleofection, transfection, and lentiviral transduction were compared for optimal gene delivery to NP cells. Transduction was most efficient in terms of transgene expression (37%), cell viability (39%), and long-term reporter expression (>3 months). Further, the constitutive gene promoters, cytomegalovirus, elongation factor 1α, and ubiquitin-C, exhibited comparable silencing (20–30%) in NP cells over a 2-month period, suggesting their suitability for long-term reporter expression studies. Transduced NP cells maintained their progenitor state and differentiation potential, as demonstrated by expression of endogenous NP markers and neuronal markers after differentiation. We also detected reporter expression in astrocytes generated from NP cells transduced with an astrocyte-specific gene promoter, glial fibrillary acidic protein, demonstrating the usefulness of this approach. The genetically manipulated NP cells described here offer great potential for live cell–tracking experiments, and a similar approach can as well be used for expression of proteins other than reporters.

Related Publication:
Sujoy K. Dhara, Brian A. Gerwe, Anirban Majumder, Mahesh C. Dodla, Nolan L. Boyd, David W. Machacek, Kowser Hasneen, Steven L. Stice. Genetic Manipulation of Neural Progenitors Derived from Human Embryonic Stem Cells. Tissue Engineering Part A. -Not available-, ahead of print. doi:10.1089/ten.tea.2009.0155

Neuromics' Stem Cell Markers Referenced:
Musashi-1, Rabbit, 1:100 , Catalog#:RA14128
Nestin, Mouse 1:500, Catalog#: MO15012
Tuj 1 (Neuron-specific class III beta-tubulin), Mouse, 1:500, Catalog#: MO15013
TH, Chicken, 1:80, Catalog#: CH23006

ENS Development Markers

This is an excellent study on the developing Enteric Nervous Stem (ENS). This system is often referred to as the "second brain".

Despite this, neurogenesis has been much less studied in the ENS than in the brain. Understanding how neurons are formed in the gut is the foundation for finding cures for ENS disorders. The key finding here is that the ability of 5-HT4 receptors to unmask a regulation of enteric neurogenesis in adult animals suggests that the mature ENS is capable of an unexpected degree of plasticity (potentially good new for discovering therapies for ENS related disorders).

Min-Tsai Liu, Yung-Hui Kuan, Jingwen Wang, René Hen, and Michael D. Gershon. 5-HT4 Receptor-Mediated Neuroprotection and Neurogenesis in the Enteric Nervous System of Adult Mice. The Journal of Neuroscience, August 5, 2009, 29(31):9683-9699; doi:10.1523/JNEUROSCI.1145-09.2009.

More details @ http://www.anxietyinsights.info/serotonin_the_gut_and_neurogenesis.htm

On a side note, this publication is rich with excellent ICC and Western Blot images of a variety of Neurogenesis Markers. This include referencing of 4 of our markers:

GFAP-CH22102-ICC Dilution 1:2000
Musashi-1-RA14128-ICC Dilution 1:100 WB Dilution 1:1000
Neurofilament NF-H-CH22104-ICC Dilution 1:2000
S100B-RA25022-ICC Dilution 1;1000