Would BD Cell Recovery Solution work for this? Or Dispase/Accutase?
Hi! I have successfully established a 3D culture of primary murine prostate epithelial cells and I would like to subculture these cells. I was wondering if anyone has a protocol and tips for this. Thank you!
It does depend on the adhesion properties of your epithelial cells especially if it's 3D vs. monolayer. I have found that Accutase tends to be quite gentle on the cells and it great for single cell dissociation. If you are after small clumps (rather than single cells) then I would recommend Stem Cell Technologies' ReLeSR which is gentler.
Why would you want to grow epithelial cells in 3D? Depending on which epithelia you are trying to model they are tissue surface barrier cells that and in vivo do not normally grow inside the ECM. So second question why Matrigel? This is tumor matrix which is variable in composition from lot to lot and not a normal ECM. A lot of people use it because it supposedly contains several growth promoters. But which ones and how much? If you want a create a true model of the epithelium then you should pair your epithelial cells with the appropriate fibroblasts. Then the fibroblasts would be grown in 3D collagen type I matrix and epithelial cells plated on the surface of this 3D culture and allowed to develop a monolayer. If you want more details send me an e-mail.
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