RosetteSep® Human Cord Blood Progenitor Cell Enrichment Cocktail

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 15026, Lot# All:28581-PIS_1_0_1.pdf
•  PRODUCT INFORMATION SHEET - 15066, Lot# All:28581-PIS_1_0_1.pdf

Educational resources:

•  VIDEO: RosetteSep™: Cell Isolation Directly From Whole Blood Without Columns or Magnets
•  VIDEO: RosetteSep®: A Guide to Isolating NK Cells Directly from Whole Blood
•  BROCHURE: One-Step Cell Enrichment Directly From Whole Blood
•  BROCHURE: Ready Sep Go – Cell Separation In As Little As 25 Minutes
•  VIDEO: RosetteSep® for HLA Analysis

MSDS:

•  15016C_15026C-MSDS.pdf

FAQS:

• 
  HOW ROSETTESEP™ WORKS
  
  
  • Q. WHAT IS ROSETTESEP™?
    A. RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.
  • Q. HOW DOES ROSETTESEP™ WORK?
    A. The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).
  • Q. WHAT FACTORS AFFECT CELL RECOVERY?
    A. The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.
  
  CELL TYPES AND SAMPLE SOURCES
  
  
  • Q. WHICH CELL SAMPLES CAN ROSETTESEP™ BE USED WITH?
    A. RosetteSep™ can be used with leukapheresis samples, bone marrow or buffy coat, as long as: the concentration of cells does not exceed 5 x 10⁷ per mL (can dilute if necessary); and there are at least 20 RBCs for every nucleated cell (RBCs can be added if necessary).
  • Q. CAN ROSETTESEP™ BE USED WITH PREVIOUSLY FROZEN OR CULTURED CELLS?
    A. Yes. Cells should be re-suspended at 2 - 5 x 10⁷ cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.
  • Q. CAN ROSETTESEP™ BE USED TO ENRICH PROGENITORS FROM CORD BLOOD?
    A. Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll™, which can lead to a higher RBC contamination than peripheral blood separations.
  • Q. DOES ROSETTESEP™ WORK WITH MOUSE CELLS?
    A. No, but we have developed EasySep™, a magnetic-based cell isolation system which works with mouse and other non-human species.
  
  ANTI-COAGULANTS
  
  
  • Q. WHICH ANTICOAGULANT SHOULD BE USED WITH ROSETTESEP™?
    A. Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.
  • Q. SHOULD THE ANTICOAGULANT BE WASHED OFF BEFORE USING ROSETTESEP™?
    A. No, the antibody cocktail can be added directly to the sample.

This product has been used in:

1. Val??rie Vanneaux et al. In vitro and in vivo analysis of endothelial progenitor cells from cryopreserved umbilical cord blood: are we ready for clinical application?Cell Transplant 19 (9) 1143-1155 (May 4, 2010)
2. Il-Kyoo Park et al. The Axl/Gas6 pathway is required for optimal cytokine signaling during human natural killer cell development.Blood 113 (11) 2470-2477 (March 12, 2009)
3. W Mitchell et al. Mapping early conformational changes in {alpha}IIb and {beta}3 during biogenesis reveals a potential mechanism for {alpha}IIb{beta}3 adopting its bent conformationBlood 109 (9) (May 1, 2007)
4. Claudia N. Renn et al. TLR Activation of Langerhans Cell-Like Dendritic Cells Triggers an Antiviral Immune ResponseJ Immunol 177 (1) 298-305 (July 1, 2006)
5. Catherine Frelin et al. Targeting NF-{kappa}B activation via pharmacologic inhibition of IKK2-induced apoptosis of human acute myeloid leukemia cellsBlood 105 (2) (January 15, 2005)
6. Robert E Hunger et al. Langerhans cells utilize CD1a and langerin to efficiently present nonpeptide antigens to T cells.J Clin Invest 113 (5) 701-708 (March 2004)
7. Oscar K Lee et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood.Blood 103 (5) 1669-1675 (March 1, 2004)
8. F Z Bischoff et al. Intact fetal cell isolation from maternal blood: improved isolation using a simple whole blood progenitor cell enrichment approach (RosetteSep).Clin Genet 63 (6) 483-489 (June 2003)

FACS Profile Results With RosetteSep® Human Cord Blood Progenitor Cell Enrichment Kit

RosetteSep® Procedure