Lymphoprep™

Density gradient medium for the isolation of mononuclear cells

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Density gradient medium for the isolation of mononuclear cells
From: 90 USD

Overview

Lymphoprep™ is a density gradient medium recommended for the isolation of mononuclear cells from peripheral blood, cord blood, and bone marrow by exploiting differences in cell density. Granulocytes and erythrocytes have a higher density than mononuclear cells and therefore sediment through the Lymphoprep™ layer during centrifugation. Lymphoprep™ can be substituted for Ficoll-Paque™ without any need to change your existing protocols and is fully compatible with both SepMate™ and RosetteSep™ .

Lymphoprep™ has a density of 1.077 g/mL.
Contains:
• Sodium diatrizoate (9.1% w/v)
• Polysaccharide (5.7% w/v)
• Other ingredients
Subtype:
Density Gradient Media
Cell Type:
Mononuclear Cells
Species:
Human
Sample Source:
Bone Marrow; Cord Blood; Whole Blood
Selection Method:
Negative
Application:
Cell Isolation
Brand:
Lymphoprep
Area of Interest:
Immunology

Scientific Resources

Product Documentation

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Educational Materials

(7)

Data and Publications

Data

Figure 1. Purity and Recovery of Cells from Whole Blood When Using Cost-Effective Lymphoprep™ is Comparable to Using Ficoll-Paque™ PLUS

(A) Density gradient centrifugation of peripheral whole blood using Lymphoprep™ results in similar cell purity of mononuclear cells including T cells, B cells, NK cells and monocytes compared to Ficoll-Paque™ PLUS. (B) The recovery of total mononuclear cells and CD45+ cells is also similar. (n = 5, Mean ± SD).

Figure 2. Purity and Recovery of Cells from Cord Blood When Using Cost-Effective Lymphoprep™ is Comparable to Using Ficoll-Paque™ PLUS

(A) Density Gradient centrifugation of cord blood using Lymphoprep™ results in similar cell purity of mononuclear cells including T cells, B cells, NK cells and monocytes compared to Ficoll-Paque™ PLUS. (B) The recovery of total mononuclear cells and CD45+ cells is also similar. (n = 4, Mean ± SD).

Publications

(27)
Blood advances 2020 oct

Acute myeloid leukemia-induced remodeling of the human bone marrow niche predicts clinical outcome.

Y. Chen et al.

Abstract

Murine models of myeloid neoplasia show how leukemia infiltration alters the hematopoietic stem cell (HSC) niche to reinforce malignancy at the expense of healthy hematopoiesis. However, little is known about the bone marrow architecture in humans and its impact on clinical outcome. Here, we dissect the bone marrow niche in patients with acute myeloid leukemia (AML) at first diagnosis. We combined immunohistochemical stainings with global gene expression analyses from these AML patients and correlated them with clinical features. Mesenchymal stem and progenitor cells (MSPCs) lost quiescence and significantly expanded in the bone marrow of AML patients. Strikingly, their HSC- and niche-regulating capacities were impaired with significant inhibition of osteogenesis and bone formation in a cell contact-dependent manner through inhibition of cytoplasmic $\beta$-catenin. Assessment of bone metabolism by quantifying peripheral blood osteocalcin levels revealed 30{\%} lower expression in AML patients at first diagnosis than in non-leukemic donors. Furthermore, patients with osteocalcin levels ≤11 ng/mL showed inferior overall survival with a 1-year survival rate of 38.7{\%} whereas patients with higher osteocalcin levels reached a survival rate of 66.8{\%}. These novel insights into the human AML bone marrow microenvironment help translate findings from preclinical models and detect new targets which might pave the way for niche-targeted therapies in AML patients.
Science advances 2020 may

Efficient blockade of locally reciprocated tumor-macrophage signaling using a TAM-avid nanotherapy.

S. J. Wang et al.

Abstract

Interpreting how multicellular interactions in the tumor affect resistance pathways to BRAF and MEK1/2 MAPK inhibitors (MAPKi) remains a challenge. To investigate this, we profiled global ligand-receptor interactions among tumor and stromal/immune cells from biopsies of MAPK-driven disease. MAPKi increased tumor-associated macrophages (TAMs) in some patients, which correlated with poor clinical response, and MAPKi coamplified bidirectional tumor-TAM signaling via receptor tyrosine kinases (RTKs) including AXL, MERTK, and their ligand GAS6. In xenograft tumors, intravital microscopy simultaneously monitored in situ single-cell activities of multiple kinases downstream of RTKs, revealing MAPKi increased TAMs and enhanced bypass signaling in TAM-proximal tumor cells. As a proof-of-principle strategy to block this signaling, we developed a multi-RTK kinase inhibitor nanoformulation that accumulated in TAMs and delayed disease progression. Thus, bypass signaling can reciprocally amplify across nearby cell types, offering new opportunities for therapeutic design.
Scientific reports 2020 jun

A Simple and Scalable Strategy for Analysis of Endogenous Protein Dynamics.

M. K. Schwinn et al.

Abstract

The ability to analyze protein function in a native context is central to understanding cellular physiology. This study explores whether tagging endogenous proteins with a reporter is a scalable strategy for generating cell models that accurately quantitate protein dynamics. Specifically, it investigates whether CRISPR-mediated integration of the HiBiT luminescent peptide tag can easily be accomplished on a large-scale and whether integrated reporter faithfully represents target biology. For this purpose, a large set of proteins representing diverse structures and functions, some of which are known or potential drug targets, were targeted for tagging with HiBiT in multiple cell lines. Successful insertion was detected for 86{\%} of the targets, as determined by luminescence-based plate assays, blotting, and imaging. In order to determine whether endogenously tagged proteins yield more representative models, cells expressing HiBiT protein fusions either from endogenous loci or plasmids were directly compared in functional assays. In the tested cases, only the edited lines were capable of accurately reproducing the anticipated biology. This study provides evidence that cell lines expressing HiBiT fusions from endogenous loci can be rapidly generated for many different proteins and that these cellular models provide insight into protein function that may be unobtainable using overexpression-based approaches.
Science advances 2020 jun

A highly potent lymphatic system-targeting nanoparticle cyclosporine prevents glomerulonephritis in mouse model of lupus.

R. Ganugula et al.

Abstract

Cyclosporine A (CsA) is a powerful immunosuppressant, but it is an ineffective stand-alone treatment for systemic lupus erythematosus (SLE) due to poor target tissue distribution and renal toxicity. We hypothesized that CD71 (transferrin receptor 1)-directed delivery of CsA to the lymphatic system would improve SLE outcomes in a murine model. We synthesized biodegradable, ligand-conjugated nanoparticles [P2Ns-gambogic acid (GA)] targeting CD71. GA conjugation substantially increased nanoparticle association with CD3+ or CD20+ lymphocytes and with intestinal lymphoid tissues. In orally dosed MRL-lpr mice, P2Ns-GA-encapsulated CsA increased lymphatic drug delivery 4- to 18-fold over the ligand-free formulation and a commercial CsA capsule, respectively. Improved lymphatic bioavailability of CsA was paralleled by normalization of anti-double-stranded DNA immunoglobulin G titer, plasma cytokines, and glomerulonephritis. Thus, this study demonstrates the translational potential of nanoparticles that enhance the targeting of lymphatic tissues, transforming CsA into a potent single therapeutic for SLE.
Cell reports 2020 feb

Nef-Mediated CD3-TCR Downmodulation Dampens Acute Inflammation and Promotes SIV Immune Evasion.

S. Joas et al.

Abstract

The inability of Nef to downmodulate the CD3-T cell receptor (TCR) complex distinguishes HIV-1 from other primate lentiviruses and may contribute to its high virulence. However, the role of this Nef function in virus-mediated immune activation and pathogenicity remains speculative. Here, we selectively disrupted this Nef activity in SIVmac239 and analyzed the consequences for the virological, immunological, and clinical outcome of infection in rhesus macaques. The inability to downmodulate CD3-TCR does not impair viral replication during acute infection but is associated with increased immune activation and antiviral gene expression. Subsequent early reversion in three of six animals suggests strong selective pressure for this Nef function and is associated with high viral loads and progression to simian AIDS. In the absence of reversions, however, viral replication and the clinical course of infection are attenuated. Thus, Nef-mediated downmodulation of CD3 dampens the inflammatory response to simian immunodeficiency virus (SIV) infection and seems critical for efficient viral immune evasion.
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