NeuroCult® NCFC Assay Kit (Mouse)

Recommended for:	• Quantification of neural colonies derived from neural stem and progenitor cells in primary embryonic or adult mouse CNS tissue or cultured neurospheres • Discrimination between neural stem and progenitor cells
Components:	• NeuroCult® NCFC Serum-Free Medium without Cytokines, 50 mL (Catalog #05720) • NeuroCult® NS-A Proliferation Supplement (Mouse), 50 mL (Catalog #05701) • NeuroCult® NS-A Basal Medium (Mouse), 450 mL (Catalog #05700) • Collagen Solution, 35 mL (Catalog #04902) • NeuroCult® Collagenase Solution, 5 mL (Catalog #07914) • 35 mm Culture Dishes, 70 dishes (Catalog #27100) • Gridded Scoring Dishes, 5 dishes (Catalog #27500) • Technical Manual (Manual Catalog #28709)
Accessory Products:	• EGF, Human, Recombinant (Catalog #02633) • bFGF, Human, Recombinant (Catalog #02634) • Heparin (Catalog #07980)
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Legal Statement:	* Sold under license from StemCells California, Inc. US Patent Nos. 5,750,376; 5,851,832; 5,980,885; 5,968,829; 5,981,165; 6,071,889; 6,093,531; 6,103,530; 6,165,783; 6,238,922.
Product Type:	Specialized cell culture media
Application:	Colony assays & quantification
Area of Interest:	Cancer, Neuroscience, Stem cell biology
Cell Type:	Neural stem & progenitor cells
Medium Type:	Collagen-based
Popular Product Line:	NeuroCult
Species:	Mouse

Procedures and instruction manuals:

•  MANUAL: Identification and Enumeration of Mouse Neural Stem and Progenitor Cells With the NeuroCult® Neural Colony-Forming Cell (NCFC) Assay Kit
•  PRODUCT INFORMATION SHEET - 05740, Lot# All:29577-PIS_1_1_0.pdf

Educational resources:

•  WEBINAR: Standardized Media and Reagents for Neural Stem Cell Research with NeuroCult®
•  BROCHURE: Neural Stem Cells: Standardized Media and Reagents
•  SCIENTIFIC POSTER: A novel culture assay that distinguishes between neural stem and progenitor cells
•  SCIENTIFIC POSTER: In vivo administration of EGF directly into the adult mouse subventricular zone has a greater effect on the frequency of neural progenitors than neural stem cells
•  SCIENTIFIC POSTER: Colony assay for neural stem and progenitors adapted to 24- and 96-well plate formats
•  SCIENTIFIC POSTER: An alternate method for measuring sphere-forming cell numbers to the neurosphere assay
•  MINI REVIEW: Mini-review: Neural stem cells (Catalog #29019)
•  BROCHURE: NeuroCult™ Reagents for Brain Tumor Stem Cell Research
•  WEBINAR: Neural Stem Cell Culture Systems - Neurosphere vs. Adherent Monolayer Culture
•  WEBINAR: Identification and Characterization of Neural Stem Cells: Why is This so Darned Difficult?

MSDS:

•  05720-MSDS.pdf

This product has been used in:

1. Stefano Pluchino et al. Persistent inflammation alters the function of the endogenous brain stem cell compartment.Brain 131 (Pt 10) 2564-2578 (October 2008)
2. Mohammad G Golmohammadi et al. Comparative analysis of the frequency and distribution of stem and progenitor cells in the adult mouse brain.Stem Cells 26 (4) 979-987 (April 2008)
3. Sharon A Louis et al. Enumeration of neural stem and progenitor cells in the neural colony-forming cell assay.Stem Cells 26 (4) 988-996 (April 2008)
4. Karine Sii-Felice et al. Fanconi DNA repair pathway is required for survival and long-term maintenance of neural progenitors.EMBO J 27 (5) 770-781 (March 5, 2008)
5. Kaylene M Young et al. Subventricular zone stem cells are heterogeneous with respect to their embryonic origins and neurogenic fates in the adult olfactory bulb.J Neurosci 27 (31) 8286-8296 (August 1, 2007)
6. Natalie D Bull et al. The adult mouse hippocampal progenitor is neurogenic but not a stem cell.J Neurosci 25 (47) 10815-10821 (November 23, 2005)

Background References:

1. B A Reynolds et al. Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell.Dev Biol 175 (1) 1-13 (April 10, 1996)

Product Name	Description	Catalog #
NeuroCult® Proliferation Kit (Mouse)	Medium for Expansion of Mouse Neural Stem and Progenitor Cells	05702
NeuroCult® Differentiation Kit (Mouse)	Medium for Differentiation of Mouse Neural Stem and Progenitor Cells	05704
NeuroCult® Chemical Dissociation Kit (Mouse)	Kit for Chemical Dissociation of Mouse Neurospheres	05707
NeuroCult® Enzymatic Dissociation Kit for Adult CNS Tissue (Mouse and Rat)	Kit for Enzymatic Dissociation of Adult Mouse and Rat CNS Tissue	05715
ALDEFLUOR™ Kit	ALDEFLUOR™ Kit for the Identification, Evaluation and Isolation of Stem and Progenitor Cells Expressing High Levels of ALDH	01700
EGF, Human, Recombinant	Epidermal Growth Factor	02633
0.2% Heparin Sodium Salt in PBS	0.2% Heparin Sodium Salt in Phosphate Buffered Saline	07980
NeuroCult® NCFC Assay Kit (Rat)	Assay to Quantify Neural Stem and Progenitor Cells	05742
bFGF, Human, Recombinant	Basic Fibroblast Growth Factor (bFGF)	02634

Overview of the NeuroCult® NCFC assay procedure.

Proliferative potential of cells obtained from colonies >2 mm in diameter

Cells isolated from six representative colonies (L1, L2, L3, L4, L5, L6) >2 mm in diameter were passaged in neurosphere cultures and showed high fold expansion and self-renewal ability. Cells from a colony (L7) 1 - 2 mm in diameter failed to form neurospheres past passage 3.

Cells isolated from colonies >2 mm in diameter maintain multi-lineage potential

A. Neurons (red) were detected with a mouse monoclonal ß-Tubulin III antibody. B. Astrocytes (green) were detected with a rabbit polyclonal GFAP antibody. C. Oligodendrocytes (green) were detected with a rabbit monoclonal O4 Oligodendrocyte Marker antibody.