Microencapsulation of cells in hydrogels using electrostatic potentials

• US 8,202,701 B2
• Filed: 10/07/2005
• Issued: 06/19/2012
• Est. Priority Date: 10/08/2004
• Status: Expired due to Fees

• Current Assignee: Georgia Tech Research Corporation (University System of Georgia)
• Original Assignee: Georgia Tech Research Corporation (University System of Georgia)
• Inventors: Boyan, Barbara Dale, Schwartz, Zvi, Kinney, Ramsey Christian
• Primary Examiner(s): Ford, Allison
• Assistant Examiner(s): FERNANDEZ, SUSAN EMILY
• Application Number:
  US11/576,542
• Publication Number:
  US 20080031962A1
• Time in Patent Office: 2,447 Days
• Field of Search: 435/29, 435/174, 435/182, 435/382
• US Class Current: 435/29
• CPC Class Codes: A61K 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 2533/74 Alginate
• CPC Class Codes: C12N 5/0012 Cell encapsulation
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage

Abstract

Compositions and methods for producing encapsulated cells having an average diameter of less than about 200 μm are provided. Methods for using the disclosed encapsulated cells are also provided.

Claims

1. A method for selecting a compound comprising:

(a) contacting encapsulated cells with at least one test compound, wherein the encapsulated cells are spotted on addressable locations on a substrate and the cells are encapsulated in a structure having a diameter of less than about 200 μ

m; and

(b) selecting the compound that induces or promotes a change in phenotype of the encapsulated cells contacted with the test compound compared to a control, wherein the encapsulated cells are produced by a process comprising applying an electrostatic potential to a droplet of cells suspended in a first solution comprising one or more types of monomers, wherein the electrostatic potential is in an amount sufficient to disrupt the surface tension of the droplet, and dropping the droplet into a polymerization solution from a distance sufficient to produce a structure encapsulating from about 60 to about 100 cells with a diameter of less than about 200 μ

m, wherein the polymerization solution comprises a nutrient osmolyte and an ionic polymerizing agent that promotes the polymerization of the one or more types of monomers.

2. The method of claim 1, wherein the encapsulated cells comprise mammalian cells.

3. The method of claim 2, wherein the mammalian cells comprise differentiated mesenchymal cells, epithelial cells, neural cells, endothelial cells, epithelial cells, myoblasts, chondrocytes, myoblasts, osteoblasts, osteoclasts, bone marrow cells, adult stem cells, embryonic stem cells, umbilical cord blood cells, fibroblasts, or a combination thereof.

4. The method of claim 1, wherein the encapsulated cells comprise at least one exogenous nucleic acid.

5. The method of claim 1, wherein the cells are encapsulated in a polysaccharide polymer.

6. The method of claim 5, wherein the polymer comprises alginate, agarose, polyaminoacids, or a combination thereof.

7. The method of claim 1, wherein the phenotype change comprises a change in protein synthesis, DNA synthesis, gene expression, color, morphology, size, cell number, cell viability, secretion, or a combination thereof.

8. The method of claim 1, wherein the process of producing the encapsulated cells further comprises the step of treating the encapsulated cells with a second polymeric material comprising polyamino acids.

9. The method of claim 8, wherein the polyamino acids are selected from the group consisting of poly-L-lysine, poly-L-ornithine, poly-L-arginine, poly-L-asparagine, poly-L-aspartic acid, poly-benzyl-L-aspartate, poly-S-benzyl-L-cysteine, poly-γ

-benzyl-L-glutamate, poly-S-CBZ-L-cysteine, poly-ε

-CBZ-D-lysine, poly-δ

-CBZ-DL-ornithine, poly-O-CBZ-L-serine, poly-O-CBZ-D-tyrosine, poly(γ

-ethyl-L-glutamate), poly-D-glutamic acid, polyglycine, poly-γ

-N-hexyl L-glutamate, poly-L-histidine, poly(α

,β

-[N-(2-hydroxyethyl)-DL-aspartamide]), poly-L-hydroxyproline, poly(α

,β

-[N-(3-hydroxypropyl)-DL-aspartamide]), poly-L-isoleucine, poly-L-leucine, poly-D-lysine, poly-L-phenylalanine, poly-L-proline, poly-L-serine, poly-L-threonine, poly-DL-tryptophan, poly-D-tyrosine, and a combination thereof.

10. The method of claim 1, wherein the ionic polymerizing agent comprises a divalent cation.

11. The method of claim 10, wherein the divalent cation is a calcium cation.

12. The method of claim 1, wherein the droplet of cells comprises about 60 to about 100 cells suspended in a solution of alginate.

13. The method of claim 1, wherein the polymerization solution comprises at least 20 mM CaCl₂.

14. The method of claim 1, wherein the nutrient osmolyte comprises glucose.