Mouse Bone Marrow Isolation - How to increase yield of BM-derived cells? (Feb/10/2011 )
I'm having trouble isolating cells from the bone marrow of mice. Basically, I think my problem is with cutting the tissue around the leg bones. I use basic scissors to make a small puncture in the leg, and then come in with forceps to tear away the bones. I then cut at the heal and hip with the scissors, followed by more removal of tissue using the forceps. I then stick a needle in the bone to expunge the marrow (25 gauge). Perhaps I should change needle size? Any advice is appreciated.
The 25-gauge needle may be a bit small, and damage your cells. Roche recommends tissue disruption using a 0.6 mm diameter needle (21 G), where a 25 G is 0.5 mm. Aspiration could exert enough pressure to rupture your cells.