Neurosphere Dissociation Kits

Gentle and reliable dissociation
Quick procedure with high cell yield
Optimized kit with detailed protocol

Overview

The Neurosphere Dissociation Kits have been developed for the gentle and efficient generation of single-cell suspensions from cultured neurospheres. In less than an hour, the gentle enzymatic dissociation procedure efficiently yields high numbers of viable cells.
Dissociated cells are immediately ready for further in vitro or in vivo applications such as cell culture and expansion, flow cytometry, fluorescence microscopy, and molecular applications, or for cell separations using MACS® Technology .

Details

Background information
Neurospheres are non-adherent spherical clusters of cells formed when neural stem cells are cultured under appropriate conditions. Effective dissociation is important for effective pasaging and subsequent downstream applications

Some antigen epitopes are degraded by papain, others by trypsin. The kit based on papain is best suited for use with Anti-Prominin-1 MicroBeads, CD11b (Microglia) MicroBeads and Anti-A2B5 MicroBeads, whereas, for example, Anti-GLAST (ACSA-1) MicroBead Kit and Anti-PSA-NCAM MicroBeads require the use of the trypsin-based kit.
Please see Figure 2 for further information with regard to epitope sensitivity and kit selection.

Further information

Neuroscience research brochure
[PDF; 1,7 MB]

Figure 1

Mouse whole brain was dissociated using the Neural Tissue Dissociation Kit (P) and the gentleMACS™ Dissociator. Neural stem cells (NSCs) were isolated via Anti-Prominin-1 MicroBeads and subsequently cultivated in MACS® Neuro Medium supplemented with MACS Supplement B27 PLUS, penicillin/streptomycin, 2 mM L-glutamine, 20 ng/mL human EGF and FGF-2. A) Primary neurospheres formed by the NSCs after one week in culture. B) Single-cell suspension after dissociation of primary neurospheres using the Neurosphere Dissociation Kit (P). C) Secondary neurospheres formed one week after dissociation.

Figure 2

Antigen compatibility of Dissociation Kits.
For many applications, including MACS Cell Separation, it is crucial to conserve the integrity of particular cell surface epitopes during tissue dissociation. The table shows which kits conserve which specific epitopes. Epitope sensitivities have been tested using Miltenyi Biotec's antibodies if available.