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Intracellular delivery

A cell and cell membrane technology, applied in the field of intracellular delivery, can solve problems such as difficulty in developing methods and high specificity, achieve accurate and large-scale delivery, and reduce experimental noise

Active Publication Date: 2014-08-13
MASSACHUSETTS INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Existing methods are often difficult to develop and highly specific to their particular application
Therefore, existing technologies do not adequately address the problems with many clinically important cell types such as stem cells and immune cells

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0137] Example 1 - Delivery of Functionally Engineered Nanoparticles

[0138] Engineered nanoparticles have great potential as live cell imaging tools, therapeutic molecule delivery agents or even as a method of manipulating living cells by using external means such as optical or magnetic fields (Howarth, M., et al. Monovalent, reduced-size quantum dots for imaging receptors on living cells (monovalent small-size quantum dots for imaging receptors in living cells). Nature Methods 5, 397-399 (2008)). However, many of these potential applications require delivery of nanomaterials into the cytoplasm of cells. Most nanoparticles, such as QDs, need to be passivated by polymers to enable the nanoparticles to dissolve in aqueous media, which often hinders the passive diffusion of nanoparticles across cell membranes.

[0139] Microinjection of nanoparticles was considered unfeasible due to lack of specialized instruments and low throughput while electroporation enables intracellula...

Embodiment 2

[0162] Example 2 - Delivery of macromolecules

[0163] Intracellular delivery of macromolecules is a critical step for both therapeutic and research applications. Nanoparticle-mediated DNA and RNA delivery can be useful, for example, in gene therapy, while protein delivery can be used to affect cellular function both in the clinic and in the laboratory. Other materials, such as small molecules, quantum dots, or gold nanoparticles, can be delivered into the fluid of cells for applications including cancer therapy, intracellular labeling, and single-cell tracking.

[0164] To illustrate the versatility of the technique, model dextran molecules were delivered into several cell types: DC2.4 dendritic cells, neonatal human foreskin fibroblasts (NuFF) and mouse embryonic stem cells (mESC), the delivery efficiencies obtained Respectively as high as 55%, 65% and 30%. Initial implementations also showed successful delivery in primary lymphocytes, macrophages, and dendritic cells fr...

Embodiment 3

[0201] Example 3 - Stem Cells and Immune Cells

[0202] Proteins, nanoparticles, siRNA, DNA and carbon nanotubes were successfully delivered to 11 different cell types, including embryonic stem cells and immune cells. Indeed, the devices and methods of the present invention are able to deliver structurally diverse materials and their availability into primary cells that are difficult to transfect, suggesting broad research and clinical applications of the present methods.

[0203] In Example 3, each device consisted of 45 identical microfluidic channels arranged in parallel, including one or more constrictions, etched on a silicon chip and sealed with a layer of borosilicate glass. The width and length of each constriction (described in more detail below) are 4-8 microns and 10-40 microns, respectively. The devices of Example 3 were typically operated at a throughput rate of 20,000 cells / second, each device was capable of producing close to one million processed cells befor...

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Abstract

A microfluidic system for causing perturbations in a cell membrane, the system including a microfluidic channel defining a lumen and being configured such that a cell suspended in a buffer can pass therethrough, wherein the microfluidic channel includes a cell-deforming constriction, wherein a diameter of the constriction is a function of the diameter of the cell.

Description

[0001] related application [0002] This application claims priority to U.S. Provisional Application No. 61 / 548,013, filed October 17, 2011, and U.S. Provisional Application No. 61 / 684,301, filed August 17, 2012, the contents of each of which are adopted This reference is hereby incorporated herein. [0003] Federally Sponsored Research Statement [0004] This invention was made, at least in part, with Government support under Grant 5RC1 EB011187-02 awarded by the National Institutes of Health. The government has certain rights in this invention. Background technique [0005] Many pharmaceutical companies focus primarily on the development of small molecule drugs. [0006] The so-called small molecule drugs are due to the relatively small molecular size of these drugs, which allows them to diffuse freely in the body to reach their targets. These molecules can also slide across otherwise impermeable cell membranes that act as great barriers. However, a new generation of pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M3/02C12N15/89
CPCC12N15/87C12N5/0602C12M35/04B82Y5/00C12M23/16C12M35/02C12M35/00C12N5/06C12M1/02
Inventor A·沙瑞A·亚当姆R·兰格K·F·延森
Owner MASSACHUSETTS INST OF TECH
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