RosetteSep Bone Marrow Enrichment, 2 mL

This STEMCELL Technologies cell isolation cocktail enriches untouched bone marrow progenitor cells from human whole bone marrow by negative selection. The RosetteSep™ antibody cocktail crosslinks unwanted (non-progenitor) cells in human whole bone marrow to multiple red blood cells (RBCs), forming immunorosettes. These immunorosettes have increased density and pellet along with free RBCs when centrifuged over a density gradient medium, while the unlabeled desired cells are collected at the plasma-density gradient interface as a highly enriched population that is immediately available for downstream applications such as flow cytometry, cell culture, or DNA/RNA extraction.

The cocktail is supplied as a combination of monoclonal antibodies in PBS. According to the protocol, 50 µL of cocktail is added per mL of bone marrow sample, mixed, and incubated at room temperature for 20 minutes. The sample is then diluted, layered over a density gradient medium of 1.077 g/mL (such as Lymphoprep™), and centrifuged at 1200 x g for 20 minutes with the brake off. Enriched cells are washed at 300 x g for 10 minutes. The starting nucleated cell concentration should not exceed 5 x 10^7 cells/mL. The cocktail is stored at 2 - 8°C and should not be frozen.

Reported performance includes a typical bone marrow progenitor cell enrichment of 25 ± 10-fold. In a representative example, the purities of the starting and final isolated fractions were 0.6% and 17%, respectively (29-fold enrichment of CD34+ cells). Manufacturer figures also indicate a CD34+ cell enrichment of 25 ± 10% and a fold enrichment of colony-forming cells of 10–53 fold. Purity can be assessed using fluorochrome-conjugated antibodies such as Anti-Human CD34, Clone 581 or Clone 8G12. The product is for research use only and not intended for human or animal diagnostic or therapeutic uses.