Hello. I am trying to perform cell dissociation for the whole adult mouse brains (pnd90). I used Worthington Papain Dissociation System for a while, but after dissociation, cell viability was too low and the kit is a little too expensive for me to purchase it constantly. Does anyone have a cheaper protocol using media such as Hibernate, and enzymes like Papain with Dnase etc.? How much papain and Dnase would be enough to dissociate cells of a whole adult mouse brain and how long should papain incubation be in this case, according to your experiences? Thanks in advance.
Hi,
Are you trying to culture the adult dissociated cells? The viability is always going to be very poor with adult neuron dissociation.
Miltenyi make dissociation kits for postnatal neurons which are much cheaper than worthington (Plus I think worthington is optimised for early postnatal). The following is another of their kits made for adult brain dissociation, it is expensive but does many preparations:
https://www.miltenyibiotec.com/GB-en/products/macs-sample-preparation/tissue-dissociation-kits/adult-brain-dissociation-kit-mouse-and-rat.html
Let me know if you find a solution!
Sorry for answering so late. I was so hopeless and stopped checking my question status. Thank you so much, I will try this and let you know the outcome!
- Badges
- Science method