Selective delivery of material to cells

A technology of cells and blood cells, applied in the field of selective delivery of materials to cells

Active Publication Date: 2016-08-10
MASSACHUSETTS INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing technologies that rely on nanoparticles, electric fields, pore-forming chemistry, etc. are able to deliver some materials into certain cell types, but often target the physical properties of the target cell in a completely indifferent manner

Method used

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  • Selective delivery of material to cells
  • Selective delivery of material to cells
  • Selective delivery of material to cells

Examples

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Embodiment 1

[0045] Whole blood or other cell suspensions are also processed with unlabeled and / or antibody-coated magnetic beads. These cells are then isolated using a high-fidelity, magnetic enrichment system for rare cells. A nanopore technology can also be used for high-purity isolation by imaging and automatically retrieving the desired individual cells from an elastic array of 84,672 subnanometer-sized pores.

[0046] Obtaining single, viable, pure, intact CTCs of different phenotypes allows characterization of the host from genomic to functional level efforts with immediate clinical and metastatic relevance. This method allows a highly sensitive and specific enrichment of live, distinct CTCs with reduced bias.

Embodiment 2

[0048] Magnetic nanoparticles were delivered to tumor cell lines & PBMCs. Nanoparticle delivery to EpCAM-expressing, epithelial cancer cell lines, such as HT-29, LNCaP, and SK-BR-3, was compared with bulk peripheral blood mononuclear cell (PBMC) suspensions derived from human blood.

[0049] 10 nm iron oxide nanoparticles with a polyethylene glycol (PEG) surface coating were delivered into cancer cells mixed with whole blood, and the resulting target cell mixture was processed by using the cell separation device described above. For example, microfluidic delivery systems have been used to induce rapid mechanical deformation of cells to create transient voids in cell membranes (attached figure 1 ). The method has demonstrated the ability to deliver a range of materials, including proteins, RNA, DNA, and nanoparticles, to a variety of cell types, and is applicable to whole blood, a medium that typically poses problems for microfluidic systems.

[0050] Exemplary marker molecul...

Embodiment 3

[0053] A combined immunological and morphological approach was performed as follows. After processing through the device based on cell size, the cells are treated with an antibody or other tumor cell specific ligand such as a fluorescently labeled anti-CD45 antibody. The sensitivity and specificity of three different isolation methods were compared: 1) device only, 2) anti-CD45 antibody only, 3) device + anti-CD45 antibody. Morphological markers (device) + immunolabeling (e.g. anti-CD45 antibody) were found to exhibit superior sensitivity relative to the other two techniques used alone (attached figure 2 ). For example, a 2-5 fold increase in sensitivity and / or a 2-5 fold increase in specificity compared to anti-CD45 antibody alone can be observed. Enrichment indices of more than one order of magnitude were observed (attached figure 2 ).

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Abstract

Isolating or identifying a cell based on a physical property of said cell can include providing a cell suspension; passing said suspension through a microfluidic channel that includes a constriction; passing the cell suspension through the constriction; and, contacting said cell suspension solution with a compound. The constriction can be sized to preferentially deform a relatively larger cell compared to a relatively smaller cell.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Patent Application No. 61 / 866,972, filed August 16, 2013, which is hereby incorporated by reference in its entirety. [0003] governmental support [0004] The present invention obtained R01GM101420-01A1 awarded by the National Institutes of Health, RC1EB011187-02 awarded by the National Institutes of Health, P30-CA-14051 awarded by the National Cancer Institute, and the main fund number of GRFP awarded by the National Science Foundation is Government support for #1122374. This government agency has certain rights in this invention. technical field [0005] The field of the invention relates to the size selective delivery of materials to cells. Background of the invention [0006] Intracellular delivery of materials is a challenge. Existing technologies relying on nanoparticles, electric fields, pore-forming chemistry, etc. are able to deliver some materials int...

Claims

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Application Information

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IPC IPC(8): B07B1/00
CPCG01N1/30G01N2015/1006G01N15/1459B82Y30/00C12N5/0634B01L3/502761C12Q1/04B01L2300/08C12N5/0693G01N15/1484B01L2400/0487G01N33/574B01L2200/0652G01N2015/1028G01N15/01C12M23/16
Inventor A·R·沙瑞V·A·阿达尔斯泰因松N·曹R·S·兰格J·C·洛夫K·F·延森
Owner MASSACHUSETTS INST OF TECH
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