Figure 2.

Preparation of the flush/crush and bone chips fractions from the bone marrow biopsy

(A) A cleaned mortar/pestle and a 70 μm cell strainer is placed on a 50 mL Falcon tube.

(B) The BM biopsy is flushed with a 1 mL insulin syringe or using a P1000 pipette tip into the cleaned mortar (or directly through the strainer).

(C) BM biopsy is placed in clean mortar with 5 mL Cell Isolation buffer (2%FBS/PBS).

(D) BM biopsy is gently crushed using the pestle, gently tapping directly onto the bone fragments.

(E) Crushed cells are transferred to the 70 μm cell strainer and collected with the Flushed fraction.

(F) Remaining crushed bone fragments are transferred to a new 50 mL falcon tube and will be digested with Collagenase II enzyme.