Dissociating cells from specific tissue types requires specific parameters for tissue aggitation to obtain a high volume of viable, culturable cells. The Miltenyi gentleMACS Dissociator optimizes this task with a simple, practical protocol. In this publication the use of this apparatus on nerual tissue is explained.
The gentleMACS Dissociator facilitates the standardized preparation of single-cell suspensions from neural tissues in a closed system. Neural Tissue Dissociation Kits are optimized to preserve antigen epitopes needed for further applications like immunostainings and immunomagnetic cell separation1-5. In this protocol we show the gentle enzymatic dissociation of mouse brain using the gentleMACS Dissociator and the Neural Tissue Dissociation Kit (P), yielding in 97% viable cells. 100 mg of neural tissue yields between 5×106 and 1×107 cells, depending on the age of the host tissue. The targeted Prominin-1+ cells were isolated using Anti-Prominin-1 MicroBeads and subsequently taken into culture. The protocol is shown to be even more effective with the additional use of Myelin Removal Beads when working with neural tissue derived from mice >P7.
The authors have nothing to disclose.