Encapsulation of cells in biologic compatible scaffolds by coacervation of charged polymers

• US 8,343,742 B2
• Filed: 01/25/2010
• Issued: 01/01/2013
• Est. Priority Date: 07/26/2004
• Status: Expired due to Fees

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Agency For Science Technology and Research
• Inventors: Yu, Hanry, Toh, Yi Chin
• Primary Examiner(s): STANFIELD, CHERIE MICHELLE
• Application Number:
  US12/693,300
• Publication Number:
  US 20100136649A1
• Time in Patent Office: 1,072 Days
• Field of Search: None
• US Class Current: 435/182
• CPC Class Codes: A61L 27/165 Rubbers
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/44 having a macromolecular matrix
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: C08L 33/12 Homopolymers or copolymers of methyl methacrylate
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

This invention relates to a method for the encapsulation of cells in biologic compatible three dimensional scaffolds and the use of such cells encapsulated in a scaffold. The cells are embedded in a charged polymer that is complex coacervating with an oppositely charged polymer within biologic compatible scaffolds. The polymer complex embedding the cells is forming an ultra thin membrane on the surface of the three dimensional scaffold.

Claims

1. A method for encapsulating at least one cell in a biologically compatible scaffold comprising:

(a) providing a first substrate coated with a first charged polymer;

(b) placing a scaffold comprising at least one cell embedded in a second charged polymer having an electrical charge opposite to that of the first charged polymer onto the first charged polymer on the first substrate;

(c) placing a second substrate coated with a third charged polymer to form a sandwich structure with the first substrate and the scaffold arranged therebetween, wherein the first polymer and the third polymer are of the same electrical charge; and

d) forming by complex coacervation of the second charged polymer with the first and third charged polymers a membrane formed of a polymer complex, wherein the polymer complex encapsulates the at least one cell therein.

2. The method of claim 1, wherein an attachment-dependent cell is used as the at least one cell.

3. The method of claim 1, wherein an attachment-independent cell is used as the at least one cell.

4. The method of claim 1, wherein the charged polymers form polymer complexes being permeable to substances necessary to sustain the normal metabolic functions of the at least one cell and to products released by the at least one cell.

5. The method of claim 1, wherein the first charged polymer and the third charged polymer are identical.

6. The method of claim 1, wherein the second charged polymer is selected from the group consisting of chitosan, polyanionic alginate, negatively or positively charged collagen, carboxymethylcellulose (CMC) and polyanionic alginate.

7. The method of claim 1, wherein the first charged polymer is selected from the group consisting of terpolymer, Ca²⁺, chondroitin sulfate A-chitosan and polycationic poly(L-lysine).

8. The method of claim 6, wherein the positively charged collagen is used as said second charged polymer.

9. The method of claim 8, wherein a negatively charged terpolymer comprising at least one of acrylic acid and methacrylic acid and at least one of hydroxyethyl methacrylate and hydroxylpropyl methacrylate is used as said first charged polymer.

10. The method of claim 9, wherein a terpolymer consisting of 25% 2-hydroxyethyl methacrylate, 25% methacrylic acid and 50% methyl methacrylate (HEMA-MAA-MMA) is used as said first charged polymer.

11. The method of claim 9, wherein the degree of cross linking of the polymer complex formed by the reaction of the positively charged collagen with said terpolymer is controlled by varying the reaction time (contact time) between said polymers.

12. The method of claim 1, wherein the degree of cross linking of the polymer complex can be altered by varying the charge density of the polymers.

13. The method of claim 12, wherein a slightly methylated polymer and/or a highly methylated polymer is used, wherein the slightly methylated polymer has a charge density of between 0.9and 1.7 CE and the highly methylated polymer has a charge density of between 1.7 and 2.5 CE.

14. The method of claim 1, wherein the second substrate coated with the third charged polymer is contacted with the first substrate coated with the first charged polymer and the scaffold arranged therebetween for about 30 seconds to about 1 hour.

15. The method of claim 1, wherein the first substrate comprises a material selecting from the group consisting of glass, plastic, silicon, a metal and a metal oxide.

16. The method of claim 2, wherein the attachment-dependent cell is selected from the group consisting of a bone marrow stroma cell, a calvarial osteoblast, a Langerhans cell, a hepatocyte, a chondrocyte, a cardiac myocyte, a sinusoidal endothelial cell, a dermal fibroblast, a kerotinocyte and an oligodendrocyte.

17. The method of claim 3, wherein the attachment-independent cell is selected from the group of consisting of a hematopoetic stem cell, a T-lymphocyte, a macrophage and a neutrophil.