Massachusetts Institute of Technology

Harvard College

Stemcell Technologies Canada Inc.

Sqz Biotechnologies Company

Leland Stanford Junior University

Armon R Sharei

Klavs F Jensen

Robert Samuel Langer

Howard Bernstein

J Christopher Love

Viktor A Adalsteinsson

Maisam Dadgar

Scott Loughhead

Robert S Langer

Nahyun Cho

Darrell J Irvine

Ulrich Von Andrian

Pamela A Basto

Marc J Lajoie

Siddharth Jhunjhunwala

James Robbins Abshire

Jacquin Clarence Niles

Jacquelyn L Sikora Hanson

Shirley Mao

Sophia Liu

Jonathan B Gilbert

George Hartoularos

Matthew Booty

Megan Heimann

Kelan Hlavaty

Zhenan Bao

Andrea Adamo

Robert S Langer, Jr

Stefan Christian Bernhardt Mannsfeld

Gregory Lee Szeto

Robert Langer

Tarek Abdeljawad

Jason Murray

David Chirgwin

Harry An

Finola Moore

Xiaoyun Ding

Gregory Szeto

Bu Wang

Katarina Blagovic

Emrah Ilker Ozay

Defne Yarar

Carolyne Kelly Smith

Marc Lajoie

Tia Ditommaso

Kirubel Frew

A microfluidic chip for causing the delivery of a payload to a cell comprises a plurality of constrictions configured to allow a cell suspension to flow through one or more of the plurality of constrictions from a first fluid flow region to a second fluid flow region within the microfluidic chip, wherein a cross-sectional width of each of the plurality of constrictions is less than a diameter of cells in the cell suspension, such that membranes of the cells are perturbed when passing through the constrictions such that a payload is able to pass through the perturbed cell membranes, and wherein a quotient of a cross-sectional area over a cross-sectional perimeter of each of the plurality of constrictions is greater than greater than or equal to 0.5 μm.

High-throughput microfluidic chip having parallelized constrictions for perturbing cell membranes

The current subject matter includes methods, systems, articles, and techniques to deliver material to anucleate cells, such as red blood cells. Using a rapid deformation based microfluidic system, loading of red blood cells with macromolecules of different sizes has been shown. Although delivery to some mammalian cells, such as cancer cell lines and fibroblasts had been previously demonstrated using this technique, those designs were incompatible with RBCs that have dramatically different physical properties. Through the use of smaller constriction sizes, high speeds and different buffers successful delivery to red blood cells can be achieved. By enabling robust delivery to red blood cells in a simple, scalable manner, the current subject matter can be implemented in a diversity of applications that deliver material to study red blood cell diseases and/or use red blood cells as a therapeutic platform. Related apparatus, systems, techniques, and articles are also described.

Delivery of materials to anucleate cells

A method and device for preferentially delivering a compound such as an antigen to the cytosol of an immune cell. The method comprises passing a cell suspension comprising the target immune cell through a microfluidic device and contacting the suspension with the compound(s) or payload to be delivered.

Delivery of biomolecules to immune cells

The present invention provides methods for inducing tolerance and/or suppressing an immune response to an antigen by passing a cell suspension containing an anucleate cell through a constriction, wherein the constriction deforms the cell thereby causing a perturbation of the cell such that an antigen and/or tolerogenic factor enters the cell. In some embodiments, the anucleate cell is delivered to an individual and the antigen is delivered to and processed in a tolerogenic environment to induce tolerance and/or suppress an immune response to the antigen.

Intracellular delivery of biomolecules to induce tolerance

The present application provides formulations of modified peripheral blood mononuclear cells (PBMCs), wherein the formulation comprises: PBMCs comprising at least one antigen (e.g., a human papillomavirus (HPV) antigen) and a cryopreservation medium. In some embodiments, the PBMCs are conditioned by incubating the PBMC in the presence of an adjuvant.

Formulations of PBMCs

Gene editing can be performed by introducing gene-editing components into a cell by mechanical cell disruption. Related apparatus, systems, techniques, and articles are also described.

Gene editing through microfluidic delivery

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.

Selective delivery of material to cells

The present application provides peripheral blood mononuclear cells comprising an antigen, methods of manufacturing such PBMCs, and methods of using such PBMCs, such as for modulating an immune response in an individual. In some embodiments, the PBMCs are conditioned by incubating the PBMC in the presence of an adjuvant.

Delivery of biomolecules to PBMCs to modify an immune response

The present disclosure pertains to methods for delivering a compound into a cell comprising a cell wall, including passing a cell suspension through a constriction, wherein said constriction deforms the cell comprising a cell wall, thereby causing a perturbation of the cell such that the compound enters the cell, wherein said cell suspension is contacted with the compound.

Intracellular delivery of biomolecules to cells comprising a cell wall

Gene editing can be performed by introducing gene-editing components into a cell by mechanical cell disruption. Related apparatus, systems, techniques, and articles are also described. The methods and systems of the invention solve the problem of intracellular delivery of gene editing components and gene editing complexes to target cells. The results described herein indicate that delivery of gene editing components, e.g., protein, ribonucleic acid (RNA), and deoxyribonucleic acid (DNA), by mechanical disruption of cell membranes leads to successful gene editing. Because intracellular delivery of gene editing materials is a current challenge, the methods provide a robust mechanism to engineer target cells without the use of potentially harmful viral vectors or electric fields.

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.

Intracellular delivery

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Watertown, MA, United States of America

Armon R Sharei