RosetteSep™ DM-L Density Medium

Density gradient medium

RosetteSep™ DM-L Density Medium

Density gradient medium for use with RosetteSep™ CD3+ T cell, CD4+ T cell, CD8+ T cell, and B cell enrichment cocktails

Size 100 mL

Catalog # 15705

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Overview

RosetteSep™ DM-L is a density gradient medium designed specifically for use with RosetteSep™ cocktails for the enrichment of specific human lymphocyte populations (CD3+ T cells, CD4+ T cells, CD8+ T cells, and B cells) from whole blood. RosetteSep™ DM-L has a density of 1.081 g/mL.

This product carries the CE mark and is available as a Class I in vitro diagnostic device in the European Union and Australia.

Please Note: In light of the new EU Regulation 2017/746 (IVDR) of the European Parliament and of the Council on in vitro diagnostic medical devices, which is expected to become effective in 2022, STEMCELL has conducted a complete review of our RosetteSep™ CE-IVD products. We have determined that our current CE-marked IVD RosetteSep™ products will no longer carry the CE mark as “For In-Vitro Diagnostic Use” and will be labelled “For Research Use Only” labelling. These changes will take effect in December 2021. For more information please contact the Quality Assurance and Regulatory Department at qaschangenote@stemcell.com.

• Iodixanol
• Hetastarch
• Sodium chloride
• Sodium lactate
• Dextrose
• Calcium chloride
• Potassium chloride
• Magnesium chloride
• Water

Density Gradient Media

B Cells, T Cells

Human

Buffy Coat, Whole Blood

Negative

Cell Isolation, In Vitro Diagnostic

RosetteSep

Immunology

Scientific Resources

Product Documentation

Document Type	Product Name	Catalog #	Lot #	Language
Document Type Product Information Sheet 1	Product Name RosetteSep™ DM-L Density Medium	Catalog # 15705	Lot # All	Language English
Document Type Product Information Sheet 2	Product Name RosetteSep™ DM-L Density Medium	Catalog # 15705	Lot # All	Language Multi
Document Type Safety Data Sheet	Product Name RosetteSep™ DM-L Density Medium	Catalog # 15705	Lot # All	Language English

Educational Materials (3)

Wallchart

Human Immune Cytokines

Video

1:21

Isolate Highly Purified Cells for HLA Analysis with RosetteSep™ Immunodensity Cell Isolation

Video

2:19

Isolate Cells from Whole Blood without Columns or Magnets: RosetteSep™ Immunodensity Cell Separation

Frequently Asked Question

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.

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.

Research Area

Workflow Stages

Workflow Stages for B Cell Research

Cell Sourcing

Cell Isolation

Cell Expansion

Cell Characterization

Cell Sourcing,Default Category

Workflow Stages for Total Lymphocyte Research

Cell Sourcing

Cell Isolation

Cell Characterization

Workflow Stages for T Cell Research

Cell Sourcing

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Workflow Stages for HLA Analysis

Cell Isolation

Characterization

Data and Publications

Data

Figure 1. Minimal Granulocyte Contamination Using RosetteSep™ DM-L Density Medium

Granulocyte contamination is typically less than 1% when enriching lymphocytes from fresh whole blood using RosetteSep™. There was no significant difference in the level of granulocyte contamination when using RosetteSep™ DM-L in place of Ficoll-paque PLUS to enrich lymphocytes (N=11)

Figure 2. Rosettesep™ DM-L Improves the Recovery of Lymphocytes Enriched with Rosettesep™ from Fresh Blood

Figure 3. Cell Purity Is Equivalent or Higher for Rosettesep™ Lymphocyte Enrichment Using Rosettesep™ DM-L Instead of Ficoll-paque®

Publications (1)

Proceedings of the National Academy of Sciences of the United States of America 2011 JAN Completely phased genome sequencing through chromosome sorting. Yang H et al.

Abstract

View publication

Abstract

The two haploid genome sequences that a person inherits from the two parents represent the most fundamentally useful type of genetic information for the study of heritable diseases and the development of personalized medicine. Because of the difficulty in obtaining long-range phase information, current sequencing methods are unable to provide this information. Here, we introduce and show feasibility of a scalable approach capable of generating genomic sequences completely phased across the entire chromosome.

View All Publications

Quality Statement:

PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.

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RosetteSep™ DM-L Density Medium

Overview

Related Products

Scientific Resources

Product Documentation

Educational Materials (3)

Frequently Asked Question

What is RosetteSep™?

How does RosetteSep™ work?

What factors affect cell recovery?

Which cell samples can RosetteSep™ be used with?

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

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

Does RosetteSep™ work with mouse cells?

Which anticoagulant should be used with RosetteSep™?

Should the anticoagulant be washed off before using RosetteSep™?

Product Applications

Data and Publications

Data

Publications (1)

Abstract

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