The University of North Carolina at Chapel Hill

University of California

Cell Microsystems, Inc.

Nancy L Allbritton

Christopher E Sims

Yuli Wang

Mark Bachman

Guann-Pyng Li

Pavak Kirit Shah

Bruce J Tromberg

Michael W Berns

Tatiana B Krasieva

Gavin D Meredith

Raehyun Kim

Steven C Gebhart

Peter Joseph Attayek

Nicholas C Trotta

Eric J Stanbridge

Scott Magness

Chao L Lee

Jeng-Hao Pai

Robert W McClellan

Scott Bultman

Paul Michael Armistead

Nicholas C Dobes

Chris Sims

Cynthia Jensen-McMullin

Robert W Mcclellan

Embodiments of the disclosed subject matter provide an automated method and system to isolate and collect cells using computerized analysis of images of cells and their surroundings obtained from a digital imaging device or system. Embodiments of the disclosed subject matter make use of a 'microwell array,' which can comprise a formed, elastomeric grid of indentations or “wells.” Many, most, or all of the wells in a microwell array can contain a releasable, microfabricated element, which can be referred to as a “raft.” Embodiments of the disclosed subject matter provide a system and method for cell collection that includes computerized identification and collection of rafts with isolated single cells or a specific group or groups of cells, eliminating the need for continuous human identification and selection.

Automated collection of a specified number of cells

This presently disclosed subject matter relates to an in vitro cell culture comprising a cell monolayer comprising mucus producing cells and a mucus layer, and methods of making and using the same. The methods including culturing mucus producing cells on a cell support structure under conditions to establish a mucus layer on the luminal side of the cell monolayer, thereby producing a live cell construct comprising a cell monolayer comprising mucus producing cells and a mucus layer. The mucus layer can be substantially impenetrable to micro-objects, and have a thickness of about 1 micron to about 1 cm.

In vitro cell culture mucus systems

A method for producing tissue constructs comprising two or more distinct regions, each of which comprises a different cell population or lineage is described. The method involves providing a support substrate or substrate assembly comprising two or more physically distinct regions, wherein the two or more physically distinct regions of the support substrate or substrate assembly are different from each other; and depositing/positioning one or more cells on the support substrate or substrate assembly. The cells can form a continuous monolayer with at least two zones, e.g., a proliferative zone and a nonproliferative zone, that can act as in vitro intestinal models. The models are two-dimensional, thus facilitating rapid and facile imaging. Systems comprising the tissue constructs and methods of using the constructs to study the effects of pharmaceuticals, nutraceuticals, and metabolites on intestinal cells are also described.

Formation of arrays of planar intestinal crypts possessing a stem/proliferative cell compartment and differentiated cell zone

Disclosed herein are devices, apparatuses and methods for generating self-sustaining gaseous and non-gaseous gradients across a cell support structure and for culturing one or more cells or tissues under hypoxic conditions. Methods of generating one or more gas gradients across a cell support structure include providing a luminal container having a bottom wall that is a gas permeable membrane, positioning a cell support structure above the bottom wall, positioning one or more cells or tissues on the cell support structure, and generating one or more gas gradients between the bottom wall, across the cell support structure and into a luminal reservoir. An apparatus to produce hypoxic conditions for cell cultures includes a luminal container having a bottom wall, a cell support structure on the bottom wall, and a cover that sealably closes the open top, such that a hypoxic condition can be generated in the luminal reservoir.

Devices, systems and apparatuses for generating self-sustaining hypoxic conditions and gaseous and non-gaseous chemical gradients for in vitro cell culture

An apparatus for collecting or culturing cells or cell colonies includes: a common substrate formed from a flexible resilient polymeric material and having a plurality of wells formed therein; and a plurality of rigid cell carriers releasably connected to said common substrate, with said carriers arranged in the form of an array, and with each of the carriers resiliently received in one of the wells. A method of collecting or culturing cells or cell colonies with such an apparatus is carried out by depositing a liquid media carrying cells on the apparatus so that said cells settle on or adhere to said the carriers; and then (c) releasing at least one selected carrier having said cells thereon by gradual application of release energy to each carrier from the cavity in which it is received (e.g., by pushing with a probe).

Array of micromolded structures for sorting adherent cells

An array of magnetic or paramagnetic micro-elements comprised of polysilsesquioxane is described having ultra-low-autofluorescence and other optical properties to improve microscopic imaging of cells or other objects present on the array. These materials are also amenable to chemical modification allowing surface attachment of affinity capture moieties or chemical reporters for selective binding or analysis of cells, macromolecules or other targets.

Array of micro-elements for high resolution and high content imaging and sorting of cells

A microfabricated device having at least one gas-entrapping feature formed therein in a configuration that entraps air bubbles upon wetting the feature with a solvent or solution is described. The device includes a sacrificial residue in contact with the gas-entrapping feature, the dissolution of which guides the wetting of the gas-entrapping feature.

Dissolution guided wetting of structured surfaces

A method of making a live cell construct is carried out by: (a) providing a non-cellular support having a top surface and a bottom surface, (b) contacting live undifferentiated cells to the non-cellular support, and then (c) propagating a gastrointestinal epithelial cell monolayer on said top surface. In some embodiments, the live cells in the monolayer include: (i) undifferentiated cells (e.g., stem or progenitor cells); and (ii) optionally, but in some embodiments preferably, differentiated cells (e.g., enterocytes, Paneth cells, enteroendocrine cells, tuft cells, microcells, intra-epithelial lymphocytes, and/or goblet cells). Constructs formed by such methods and methods of using the same (e.g., in high through-put screening) are also described.

Methods to generate gastrointestinal epithelial tissue constructs

A micro-patterned plate composed of an array of releasable elements surrounded by a gel or solid wall and methods of manufacture of the micropatterned plate. The surface properties of walls can be tailored if needed to be repellent or attracting to proteins. The walls can also inhibit cell attachment. The walls enable cells or other materials to be localized to the tops of the releasable elements. The individual element in the array of releasable elements can be released from the array by a mechanical force.

Micro-patterned plate composed of an array of releasable elements surrounded with solid or gel walls

Growing community of inventors

Irvine, CA, United States of America

Nancy L Allbritton