RosetteSep™ Human Cord Blood Progenitor Enrichment Kit

Immunodensity negative selection cocktail

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Complete RosetteSep™ Human Cord Blood Progenitor Enrichment Kit

Includes RosetteSep™ human cord blood progenitor kit and HetaSep™

500 mL cord blood
Catalog #15276
432 USD

Required Products

Overview

The RosetteSep™ Human Cord Blood Progenitor Cell Enrichment Cocktail with HetaSep™ is designed to isolate progenitor cells from cord blood by negative selection. The HetaSep™ minimizes the required volume of the RosetteSep™ cocktail reagent by first concentrating the sample by sedimentation with HetaSep™ (Catalog #07906), a solution of hetastarch. The hematopoietic progenitors in the resultant buffy coat are then enriched using a modified RosetteSep™ protocol that uses less RosetteSep™ cocktail than is required for an unconcentrated sample. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD61, CD66b and glycophorin A on red blood cells (RBCs). When centrifuged over a density gradient medium the unwanted cells pellet along with the RBCs. The purified progenitor cells are present as a highly enriched population at the interface between the plasma and the density gradient medium.

We regret to inform our customers that we can no longer include Ficoll-Paque™ Plus with this product. For performing cell isolation by density gradient centrifugation, we recommend purchasing Lymphoprep™ (Catalog #07801), which is available in a variety of sizes and formats. As with Ficoll-Paque™, Lymphoprep™ has a density of 1.077 g/mL, and after centrifugation, mononuclear cells remain at the plasma-Lymphoprep™ interface while granulocytes and erythrocytes sediment below.
Advantages:
• Fast and easy-to-use
• Requires no special equipment or training
• Untouched, viable cells
Components:
  • RosetteSep™ Human Cord Blood Progenitor Enrichment Kit (Catalog #15276)
    • RosetteSep™ Human Hematopoietic Progenitor Cell Enrichment Cocktail, 2 mL (Catalog #15026) x 2
    • HetaSep™, 100 mL (Catalog #07906)
Subtype:
Cell Isolation Kits
Cell Type:
Hematopoietic Stem and Progenitor Cells
Species:
Human
Sample Source:
Cord Blood
Selection Method:
Negative
Application:
Cell Isolation
Brand:
RosetteSep
Area of Interest:
Immunology; Stem Cell Biology

Scientific Resources

Educational Materials

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Frequently Asked Questions

What is RosetteSep™?

RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.

How does RosetteSep™ work?

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™).

What factors affect cell recovery?

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.

Which cell samples can RosetteSep™ be used with?

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 100 RBCs for every nucleated cell (RBCs can be added if necessary).

Can RosetteSep™ be used with previously frozen or cultured cells?

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.

Can RosetteSep™ be used to enrich progenitors from cord blood?

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.

Does RosetteSep™ work with mouse cells?

No, but we have developed EasySep™, a magnetic-based cell isolation system which works with mouse and other non-human species.

Which anticoagulant should be used with RosetteSep™?

Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.

Should the anticoagulant be washed off before using RosetteSep™?

No, the antibody cocktail can be added directly to the sample.
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Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

FACS Profile Results With RosetteSep™ Human Cord Blood Progenitor CellEnrichment Kit

Figure 1. FACS Profile Results With RosetteSep™ Human Cord Blood Progenitor Cell Enrichment Kit

Starting with fresh cord blood, the enrichment of CD34+ cells is typically 29 ± 9%.

STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.