Can Human/Mouse/Rat Neural Lineage Functional Identification Kit (Catalog # SC028) be used with both embryonic and adult neural progenitor cells?

Name: Human/Mouse/Rat Neural Lineage Functional Identification Kit
Brand: R&D Systems
SKU: SC028

The Human/Mouse/Rat Neural Lineage Functional Identification Kit has been tested with embryonic neural progenitor cells. The kit should also work with adult neural progenitor cells, as adult cells have similar growth conditions.

Human/Mouse/Rat Neural Lineage Functional Identification Kit (SC028) by R&D Systems, Part of Bio-Techne

Kit Summary

To verify the multipotency of neural progenitor cells (NPCs) by in vitro functional differentiation.

Key Benefits

Confirms that the starting population is multipotent
Reduces experimental variation
Assesses the differentiation of NPCs into astrocytes, neurons, and oligodendrocytes

Why Functionally Verify Neural Progenitor Cell Multipotency In Vitro?

Neural stem cell research studies most often require weeks of cell culture before an experimental hypothesis can be tested. See Details

The ability to verify multipotency at an early stage in the experimental design provides valuable insight that may prevent weeks of wasted effort and reagents. Moreover, confirmation of multipotency helps to ensure consistency among studies and reduce unwanted experimental variability. Neural progenitor cell (NPC) multipotency is best evaluated by functionally assessing the ability of NPCs to differentiate into multiple neural lineages.

Functional verification of NPC multipotency in vitro:

Uses a specially formulated supplement to drive reproducible differentiation.
Verifies a healthy, multipotent starting NPC population to increase consistency between studies and reduce unwanted experimental variability.
Provides results in 7-10 days to save time and reagents.
Uses a monolayer culture system, which, compared to neurosphere systems, allows direct observation of NPCs and differentiated derivatives.

Kit Components

This kit contains the following reagents to drive NPC differentiation and a marker to analyze NPCs and each of the three differentiated derivatives: See Details

Neural Maintenance Supplement (10X)
Neural Differentiation Supplement
Bovine Fibronectin (100X)
NPC Marker: Goat Anti-Rat Nestin Antigen Affinity-purified Polyclonal Antibody
Astrocyte Marker: Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody
Neuron Marker: Mouse Anti-Neuron-specific beta-III Tubulin Monoclonal Antibody
Oligodendrocyte Marker: Mouse Anti-Oligodendrocyte Marker O4 Monoclonal Antibody

The quantity of neural differentiation supplement in this kit is sufficient to make 50 mL of media for NPC expansion and 100 mL of media for NPC differentiation. This is enough media for the differentiation of two 24-well plates.

Neural stem cells provide an excellent model for research focused on neural development and neurological disorders. R&D Systems offers ready-to-use primary cortical stem cells isolated from E14.5 Sprague-Dawley rats. In addition, primary mouse cortical stem cells isolated from E14.5 CD-1 mice are available. Every lot of R&D Systems Cortical Stem Cells is validated for a high level of Nestin expression and the capacity for multi-lineage differentiation into astrocytes, neurons, and oligodendrocytes. Our cortical stem cells are tested to ensure highest quality and lot to lot consistency. Both rat and mouse Cortical Stem Cells can be optimally expanded as monolayers or neurospheres.

To complement the use of primary neural stem cells, we offer a range of supportive products, including culture media which is specifically optimized for use with neural stem cells. We offer kits to promote the in vitro proliferation of neural precursors, and kits to differentiate neural stem cells into dopaminergic neurons or oligodendrocytes. In addition, kits are available which contain panels of antibodies designed to monitor the differentiation and identification of neural precursors, astrocytes, neurons, and oligodendrocytes.

Product Specifications for Human/Mouse/Rat Neural Lineage Functional Identification Kit

Species	Human, Mouse, Rat
Source	N/A

Scientific Data Examples for Human/Mouse/Rat Neural Lineage Functional Identification Kit

	Verification of Neural Progenitor Cell Multipotency. Rat neural progenitor cells were maintained in culture and differentiated towards neural lineages using the specialized media supplement supplied in the Human/Mouse/Rat Neural Lineage Functional Identification Kit (Catalog # SC028). Neural progenitor cell multipotency was functionally verified using antibodies (also supplied in the kit) to detect phenotype-specific markers for undifferentiated neural precursors and the following differentiated derivatives: astrocytes, neurons, and oligodendrocytes. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Preparation & Storage

Shipping Conditions	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage	Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, NPC multipotency is verified using the following in vitro differentiation procedure:

Culture multipotent cells of interest
Induce astrocyte, neuron, and oligodendrocyte differentiation using a media supplement
Evaluate differentiation using mature phenotype marker antibodies and fluorescent ICC

View Graphic Procedure

Reagents Provided

Reagents supplied in the Human/Mouse/Rat Neural Progenitor Cell Functional Identification Kit (Catalog # SC028):

Neural Maintenance Supplement (500X)
Neural Differentiation Supplement (100X)
Bovine Fibronectin (100X)
NPC Marker: Goat Anti-Rat Nestin Antigen Affinity-purified Polyclonal Antibody
Astrocyte Marker: Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody
Neuron Marker: Mouse Anti-Neuron-specific beta-III Tubulin Monoclonal Antibody
Oligodendrocyte Marker: Mouse Anti-Oligodendrocyte Marker O4 Monoclonal Antibody

Note: The quantity of neural differentiation supplement in this kit is sufficient to make 50 mL of media for NPC expansion and 100 mL of medium for NPC differentiation. This is enough medium for the differentiation of two 24-well plates.

Other Supplies Required

Reagents

N-2 MAX Media Supplement (Catalog # AR009)
Poly-L-Ornithine
Phosphate buffered saline (PBS)
Penicillin-Streptomycin (100X)
Bovine serum albumin (BSA)
D-MEM/F-12 (1X)

Glucose
L-Glutamine
Sodium Bicarbonate (NaHCO₃)
Trypan blue
95% Ethanol
Deionized or distilled water

Materials

Human, mouse, or rat NPCs
24-well culture plates
12 mm coverslips

15 mL centrifuge tubes
Pipettes and pipette tips
Serological pipettes

Equipment

37 °C and 5% CO₂ incubator
Centrifuge
Hemocytometer

Inverted microscope
37 °C water bath
0.2 µm filter unit, 250 mL

Procedure Overview

	Astrocyte Differentiation	Neuron Differentiation	Oligodendrocyte Differentiation
	Prepare Poly-L-Ornithine- and Fibronectin-coated coverslips.
	Plate 0.5 -1.0 x 10⁶ NPCs in media containing Neural Maintenance Supplement. Culture cells to 50% confluency.
	Replace with fresh media after 24 hours. Culture cells for 48 hours after initial plating.
	ICC detection of Nestin to identify NPCs.
Day 1	Replace media with Neural Differentiation Media.	Replace media with Neural Differentiation Media.	Replace media with Neural Differentiation Media.
Day 4	Repeat media change every 3 days.	Repeat media change every 3 days.	Repeat media change every 3 days.
Day 7-10	ICC detection of GFAP.	ICC detection of beta-III Tubulin.	ICC detection of Oligodendrocyte Marker O4.