Methods and devices for encapsulating cells

Patent metadata

Field	Value
Publication number	US-9587213-B2
Application number	US-201313749267-A
Country	US
Kind code	B2
Filing date	Jan 24, 2013
Priority date	Jan 24, 2006
Publication date	Mar 7, 2017
Grant date	Mar 7, 2017

Abstract

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.

First claim

What is claimed is:

1. A method of aggregating cells, comprising the steps of:
a) depositing a plurality of cells into an upper cell suspension seeding chamber of a cell aggregation device having a biocompatible, polymeric hydrogel substrate that includes an upper surface and a lower surface and having at least one compartment at least partially recessed into the upper surface, the compartment being composed of the upper cell suspension seeding chamber, at least one lower cell aggregation recess and a port extending there between, the upper cell suspension seeding chamber being formed and positioned to funnel cells into the lower cell aggregation recess via the port through gravitational force and the lower cell aggregation recess being composed of a cell-repellant hydrogel and being formed and positioned to coalesce the cells into a finite region of minimum gravitational energy; and
b) incubating the cells for time sufficient to allow the cells to be funneled from the upper cell seeding chamber to the lower cell aggregation recess by gravity through the port and into the finite region; and
c) incubating the cells for a time sufficient to allow the cells to aggregate in the finite region of the lower cell aggregation recess.

2. The method of claim 1 , further comprising the step of adding a selected aggregation modifying agent to the cells during the time sufficient to allow the cells to aggregate in the finite region of the lower cell aggregation recess.

3. The method of claim 1 , wherein said cells are selected from the group consisting of osteoblasts, keratinocytes, melanocytes, hepatocytes, gliacytes, pancreatic beta cells, pancreatic exocrine cells, neural stem cells, neural precursor cells, spinal cord precursor cells, nerve cells, mammary gland cells, salivary gland cells, renal glomerular endothelial cells, tubular epithelial cells, adrenocortical and adrenomedullary cells, cardiomyocytes, chondrocytes, skeletal and smooth muscle cells, fat and fat precursor cells, corneal and crystalline lens cells, embryonic retina cells, vascular cells, endothelial cells, bone marrow stromal cells, lymphocytes, smooth muscle cells, skeletal muscle cells, cardiac muscle cells, connective tissue cells, epithelial cells, nervous system cells and nonhuman embryonic stem cells, non-embryonic and hematopoietic mammalian stem cells.

4. The method of claim 3 , wherein the connective tissue cells are at least one member selected from the group consisting of a fibroblast, a monocyte, a mast cell, a granulocyte, a plasma cell, an osteoclast, an osteocyte and a chondrocyte.

5. The method of claim 3 , wherein the epithelial cells at least one member selected from the group consisting of a skin epithelial cell, a gastrointestinal epithelial cell, a urinary tract epithelial cell, a reproductive tract epithelial cell, a liver epithelial cell, a pancreas epithelial cell, and a spleen epithelial cell.

6. The method of claim 3 , wherein the nervous system cell is at least one member selected from the group consisting of a glial cell, a neuronal cell and an astrocytes.

7. The method of claim 1 , further including the step of removing the aggregated cells from the lower aggregation recess.

Assignees

• Univ Brown

Inventors

• Morgan Jeffrey
• Chai Peter
• Napolitano Anthony
• Mathiowitz Edith
• Dean Dylan

Classifications

• C12N5/0012 Primary

  Cell encapsulation · CPC title

• C12M25/16

  Particles; Beads; Granular material; Encapsulation (chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00) · CPC title

• C12M23/12

  Well or multiwell plates (C12M25/04 takes precedence) · CPC title

• C12N5/0062

  General methods for three-dimensional culture · CPC title

• C12M25/00 Primary

  Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings · CPC title