Brown University

The General Hospital Corporation

Rhode Island Hospital

Biointraface, Inc.

Jeffrey R Morgan

Martin L Yarmush

Anubhav Tripathi

William Lee

Anthony Napolitano

Dylan Dean

Toni-Marie Achilli

Blanche Ip

Joseph M Le Doux

William Robert Patterson

Edith Mathiowitz

Adam Rago

Kim Boekelheide

John D Jarrell

George D Pins

Benjamin T Wilks

Andrew Blakely

Stephanie McCalla

Susan Hall

Hui Li

John Murphy

Jacquelyn Youssef Schell

Kali L Manning

Peter Chai

John Jarrell

Described herein are in vitro quantitative methods directed to 3D extracellular matrix (ECM) organization and mechanics. The methods do not require application of an external force, anchorage, or a scaffolding material in order to culture ECM tissue constructs, and the methods enable quantitative measurements without crosstalk noise from variables attributable to external forces, anchorage, and scaffolding material.

Quantifying cell-derived changes in collagen synthesis, alignment, and mechanics in a 3D connective tissue model

This dual-compartment liver co-culture system exposes target microtissue to metabolites, allowing for evaluating toxicity using a standard multi-well plate for high throughput analysis. Discovery scientists can rapidly select lead compounds with desirable metabolic profiles, while safety scientists can determine the safety of drugs, pesticides, and environmental chemicals. This platform is allometrically scalable, so it can mimic the natural size differences between healthy organs and, with the liver, produce sufficient quantities of metabolites. The device is simple to use and designed for immediate in vitro pathology assessment using confocal microscopy, transcriptomics, and proteomics.

Multi-compartment device containing human three-dimensional (3D) microtissues for toxicity testing

The invention provides a device and a method for the assembly of an aggregation of adherent cells, particularly a three-dimensional assembly of adherent cells.

Funnel-guided microtissue stacking and manipulation

A device for assembling aggregations of adherent cells includes a gripper moveable within an assembly vessel that fixes aggregations of adherent cells at a membrane of the gripper and, by movement of the gripper, assembles aggregations of cells on a separate membrane within the vessel, thereby creating a three-dimensional assembly of aggregations of cells that fuse and can be employed in surgical procedures as a unitary tissue of adherent cells. The aggregations of cells, as assembled, can assume three-dimensional configurations distinct from any one of the component aggregations of cells assembled.

Device and method for engineering living tissues

Molds for making cell aggregation devices include upper and lower surfaces. Methods of aggregating cells include depositing cells on in a seeding chamber, incubating the cells and removing aggregated cells. Devices for encapsulating aggregates of cells include biocompatible, bio-sustainable substrates compartments and a coating of a biocompatible, bio-sustainable polymer that completely surrounds the substrate and cell-encapsulating compartments.

Methods and devices for encapsulating cells

Drugs are screened for affects on inhibiting efflux pumps and blocking gap junction communication in tumors by culturing cells to thereby form self-assembled spheroids and incubating the spheroids. Uptake of a substrate of the efflux pump and distribution of a substrate for the efflux pump within the spheroids is imaged to thereby select drugs that inhibit the efflux pump or do not block gap junction communication. Selected drugs can then be employed to treat a tumor.

Differential effects of drugs on transport in a multi-layer 3D spheroid model

The candidates are screened and then employed by administering to patients in need thereof of a drug candidate that affects heterotypic intercellular mechanotransduction. At least two types of cells are labeled with distinct intracellular fluorescent marker labels and combined to form a cell suspension and cultured to form a microtissue, such as spheroids. The cells or the spheroids are combined with a drug candidate, either before, during or after forming the spheroids. The distribution of the different cell types is compared to that of essentially the same suspension culture in the absence of the drug candidate. Alternatively, the cell power of cells cultured in a non-adherent mold that determines at least in part, the shape of microtissue formed is measured and compared with essentially the same cell suspension cultured in the same manner in the absence of a drug candidate. A patient in need thereof can be administered a drug identified as affecting heterotypic intercellular mechanotransduction. Alternatively, a macrotissue can be formed that is employed in therapeutic treatment of a patient in need thereof.

Mechanotransduction by the synergistic action of heterotypic cell interactions

Provided are assays and methods for creating proto-tissues from aggregates of cells. The invention concerns assays and methods useful in tissue engineering and reconstruction techniques, specifically in the formation of macrotissues from microtissues using microtissue pre-culture time as a controlling parameter.

Assays and methods for fusing cell aggregates to form proto-tissues

The invention is a cell aggregation device comprising a hydrogel substrate having at least one, preferably a plurality, of cell-repellant compartments recessed into the uppermost surface. Each compartment is composed of an upper cell suspension seeding chamber having an open uppermost portion and a bottom portion, and one, or more than one, lower cell aggregation recess connected to the bottom portion of the upper cell suspension seeding chamber by a port. The diameter of the port may be fully contiguous with the walls of the chambers and walls of the recesses, or the diameter of the port may be more narrow than the walls of the chamber but fully contiguous with the walls of the recesses or more narrow than both the walls of the chamber and the walls of the recesses. The upper cell suspension seeding chambers are formed and positioned to funnel the cells into the lower cell aggregation recesses through gravitational force. The aggregation recesses are formed and positioned to promote cellular aggregation by coalescing cells into a finite region of minimum gravitational energy, increasing intercellular contact and minimizing or preventing cell adherence to the substrate. A device for encapsulating aggregates of live cells is provided. The device comprises (i) a biocompatible, bio-sustainable substrate having a cell-encapsulating face composed of one or more biocompatible, bio-sustainable, spaced-apart, cell-encapsulating compartments extending therefrom and (ii) a coating layer composed of a biocompatible, bio-sustainable polymer that completely surrounds the substrate and the cell-encapsulating compartments. A method for making the device is also provided.

Cell aggregation and encapsulation device and method

A method of making a composite from a composition including a metal oxide precursor and polymer precursor by exposing the metal oxide precursor to a gas having a humidity level of about 40% to about 70% for at least about one hour.

Method of making a composite from metal oxide and polymer precursors

The invention provides new methods for purifying and concentrating viruses. The inventors have discovered that high molecular weight proteoglycans present in retroviral stocks are co-concentrated with the retroviruses, and can inhibit retroviral transduction. The new purification and concentration methods feature treatment of virus stock with an anionic polyelectrolyte and a cationic polyelectrolyte, followed by centrifugation. The new methods minimize the amount of proteoglycan co-precipitated with the infectious virus.

Selective precipitation of viruses using polyectrolytes

The invention is based on the discovery that certain membranes, which include side chains or molecular 'brushes' having, for example, tertiary amino functional groups, can be used as highly effective filters to capture viruses/virus particles from liquids without removal of proteins. New methods based on this discovery include removing viruses from liquids such as blood or plasma, removing viruses from pharmaceuticals, concentrating and/or purifying viruses, e.g., for use in gene therapy, and producing recombinant viruses in new bioreactors. The invention also includes new methods of therapy or adjunct therapy for viral infections, in which a patient's blood or plasma is filtered through the membranes to remove viruses to reduce the viral load. The invention also includes new bioreactors and viral filters containing the membranes.

Methods for removal, purification, and concentration of viruses and methods of therapy based thereupon

Selective precipitation of viruses

Methods for removal, purification, and concentration of viruses, and methods of therapy based thereupon

The invention relates to microfabricated membranes and matrices that have a highly controlled and complex three-dimensional topography. The new micro fabricated membranes and matrices can be prepared of man-made as well as natural materials, such as materials found in naturally occurring membranes, and thus can be made in the form of tissue substitutes or analogs, such as basal lamina, dermal, or skin analogs.

Growing community of inventors

Sharon, MA, United States of America

Jeffrey R Morgan