Human/Mouse/Rat Neural Lineage Functional Identification Kit

Discontinued Product

SC028 has been discontinued.
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Verification of Neural Progenitor Cell Multipotency.
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Product Details
Procedure
Citations (2)
FAQs
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Human/Mouse/Rat Neural Lineage Functional Identification Kit Summary

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.

Specifications

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.
Species
Human Mouse Rat

Product Datasheets

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Scientific Data

Immunocytochemistry Verification of Neural Progenitor Cell Multipotency. View Larger

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.

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
 
 

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

 
Prepare Poly-L-Ornithine- and Fibronectin-coated coverslips

Prepare Poly-L-Ornithine- and Fibronectin-coated coverslips.

 
 
Plate 0.5 -1.0 x 10<sup>6</sup> NPCs in media containing Neural Maintenance Supplement

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

Replace with fresh media after 24 hours.

 

Culture cells for 48 hours after initial plating.

 

 

ICC detection of Nestin to identify NPCs.

 

 
Astrocyte Differentiation
Astrocyte
Differentiation
Neuron Differentiation
Neuron
Differentiation
Oligodendrocyte Differentiation
Oligodendrocyte
Differentiation
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.

 

Citations for Human/Mouse/Rat Neural Lineage Functional Identification Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Human Dental Pulp Cells Differentiate toward Neuronal Cells and Promote Neuroregeneration in Adult Organotypic Hippocampal Slices In Vitro
    Authors: L Xiao, R Ide, C Saiki, Y Kumazawa, H Okamura
    Int J Mol Sci, 2017-08-11;18(8):.  2017-08-11
  2. A Prominin-1-Rich Pediatric Glioblastoma: Biologic Behavior Is Determined by Oxygen Tension-Modulated CD133 Expression but Not Accompanied by Underlying Molecular Profiles.
    Authors: Donovan, Laura K, Potter, Nicola E, Warr, Tracy, Pilkington, Geoffrey
    Transl Oncol, 2012-06-01;5(3):141-54.  2012-06-01

FAQs

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

    • 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.

View all Stem Cell Product FAQs

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