Microcarriers for Stem Cell Culture

• US 20120219531A1
• Filed: 11/15/2011
• Published: 08/30/2012
• Est. Priority Date: 03/17/2008
• Status: Abandoned Application

• Current Assignee: Agency For Science Technology and Research, Singapore Health Services Pte Limited (MOH Holdings Pte Limited), National University of Singapore
• Original Assignee: Agency For Science Technology and Research, KK Women's and Children's Hospital (MOH Holdings Pte Limited), National University of Singapore
• Inventors: Oh, Steve, Reuveny, Shaul, Chen, Allen, Chan, Jerry Kok Yen, Zhang, Zhiyong
• Application Number:
  US13/296,526
• Publication Number:
  US 20120219531A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61L 2430/02 for reconstruction of bones; weight-bearing implants
• CPC Class Codes: A61L 27/3834 Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: C12N 2531/00 Microcarriers
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The present application discloses a method of generating bone tissue in vivo comprising implanting cells into a human or animal at a location where bone growth is required, wherein the cells have been obtained by the in vitro suspension culture of mesenchymal stem cells attached to microcarriers.

Claims

1. A method of generating bone tissue in vivo comprising implanting cells into a human or animal at a location where bone growth is required, wherein the cells have been obtained by the in vitro suspension culture of mesenchymal stem cells attached to microcarriers.

2. The method of claim 1 wherein the in vitro suspension culture of mesenchymal stem cells attached to microcarriers comprises:

(i) attaching mesenchymal stem cells to a plurality of microcarriers to form microcarrier-mesenchymal stem cell complexes; and

(ii) culturing the microcarrier-mesnchymal stem cell complexes in suspension culture.

3. The method of claim 1 wherein the cells are mesenchymal stem cells differentiated to, or in the process of differentiating towards, the osteogenic lineage.

4. The method of claim 3, wherein the mesenchymal stem cells differentiated to, or in the process of differentiating towards, the osteogenic lineage have been obtained by inducing osteogenic differentiation of mesenchymal stem cells obtained from said suspension culture by placing the microcarrier-mesenchymal stem cell complexes under conditions which induce the differentiation of mesenchymal stem cells.

5. The method of claim 3, wherein the mesenchymal stem cells differentiated to, or in the process of differentiating towards, the osteogenic lineage have been obtained by separating mesenchymal stem cells obtained from said suspension culture from the microcarriers and culturing the separated mesenchymal stem cells in non-microcarrier culture under conditions which induce osteogenic differentiation of mesenchymal stem cells.

6. A method of generating bone tissue in vivo, the method comprising:

(i) attaching mesenchymal stem cells to a plurality of microcarriers to form microcarrier-stem cell complexes,

(ii) culturing the microcarrier-mesenchymal stem cell complexes in suspension culture,

(iii) separating cultured cells from the microcarriers,

(iv) implanting the separated cells into a human or animal at a location where bone growth is required.

7. The method of claim 6 wherein the method comprises inducing osteogenic differentiation of the cultured cells prior to implanting said cells into a human or animal.

8. The method of claim 6, wherein prior to step (iii) the method comprises placing microcarrier-mesenchymal stem cell complexes from step (ii) under conditions which induce the osteogenic differentiation of mesenchymal stem cells.

9. The method of claim 6, wherein after step (iii) the method comprises placing the separated cells under conditions which induce the osteogenic differentiation of mesenchymal stem cells.

10. A method of culturing mesenchymal stem cells in suspension culture in vitro and causing their differentiation to an osteogenic lineage, the method comprising:

(i) attaching mesenchymal stem cells to a plurality of microcarriers to form microcarrier-stem cell complexes,

(ii) culturing the microcarrier-mesenchymal stem cell complexes in suspension culture,

(iii) inducing differentiation of the mesenchymal stem cells obtained from (ii), wherein the method comprises placing the microcarrier-mesenchymal stem cell complexes under conditions which induce the osteogenic differentiation of mesenchymal stem cells.

11. Cells which have been obtained, produced or identified by a method of culturing mesenchymal stem cells in suspension culture in vitro, the method comprising:

(i) attaching mesenchymal stem cells to a plurality of microcarriers to form microcarrier-stem cell complexes; and

(ii) culturing the microcarrier-mesenchymal stem cell complexes in suspension culture.

12. The cells of claim 11 which are mesenchymal stem cells, or mesenchymal stem cells differentiated to, or in the process of differentiating towards, the osteogenic lineage.

13. Cells according to claim 12, which are mesenchymal stem cells differentiated to, or in the process of differentiating towards, the osteogenic lineage, wherein the method further comprises the step of inducing osteogenic differentiation of mesenchymal stem cells obtained from the culture by placing the microcarrier-mesenchymal stem cell complexes under conditions which induce the differentiation of mesenchymal stem cells.

14. Cells according to claim 12, which are mesenchymal stem cells differentiated to, or in the process of differentiating towards, the osteogenic lineage, wherein the method further comprises the step of separating mesenchymal stem cells obtained from the culture from the microcarriers and culturing the separated mesenchymal stem cells in non-microcarrier culture under conditions which induce osteogenic differentiation of mesenchymal stem cells.