RosetteSep Cord Blood Kit, Kit

From STEMCELL Technologies, the RosetteSep™ Human Cord Blood Progenitor Enrichment Kit enables enrichment of untouched, viable human cord blood progenitor cells directly from cord blood by negative selection. The workflow is fast and easy-to-use, requires no special equipment or training, and yields cells that are immediately available for downstream applications such as flow cytometry, cell culture, or DNA/RNA extraction.

The kit's antibody cocktail crosslinks unwanted cells in human whole blood to multiple red blood cells, forming immunorosettes that pellet along with free RBCs when centrifuged over a density gradient medium. Desired cells are never labeled with antibody and are collected as a highly enriched population at the interface between the plasma and the density gradient medium. The cocktail is stored at 2 - 8°C and must not be frozen, while the included HetaSep™ — a 6% Hetastarch solution — is stored at 15 - 25°C.

For optimal performance, whole cord blood should be used within 24 hours of collection and kept at room temperature (15 - 25°C). Recommended medium is Dulbecco's PBS containing 2% fetal bovine serum, and the density gradient medium should have a density of 1.077 g/mL. Starting with fresh cord blood, the enrichment of CD34+ cells is typically 29 ± 9%. Purity of isolated human cord blood progenitor cells can be assessed by flow cytometry using a fluorochrome-conjugated anti-human CD34 antibody.