METHOD FOR PROMOTING DIFFERENTIATION OF STEM CELL INTO INSULIN PRODUCING CELL

• US 20090311782A1
• Filed: 12/09/2008
• Published: 12/17/2009
• Est. Priority Date: 06/11/2008
• Status: Abandoned Application

• Current Assignee: Taipei Veterans General Hospital (Government of The Republic of China), HealthBanks Biotech Co., Ltd.
• Original Assignee: Taipei Veterans General Hospital (Government of The Republic of China)
• Inventors: Fu, Yu-Show, Ho, Larry Low-Tone, Hung, Shih-Chieh, Chiou, Shih-Hwa
• Application Number:
  US12/330,716
• Publication Number:
  US 20090311782A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 2506/1353 from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

A method for promoting a differentiation of stem cells into insulin producing cells is provided. The method includes steps of suspending the stem cells in a first culture medium, aggregating the stem cells to form a cell pellet, and culturing the cell pellet in a second culture medium to promote the differentiation of the stem cells of the cell pellet into the insulin producing cells.

Claims

1. A method for promoting a differentiation of progenitor cells into insulin producing cells, comprising steps of:

suspending the progenitor cells in a first culture medium;

aggregating the progenitor cells to form a cell pellet; and

culturing the cell pellet in a second culture medium to promote the differentiation of the progenitor cells of the cell pellet into the insulin producing cells.

2. A method for promoting a differentiation of stem cells into insulin producing cells, comprising steps of:

suspending the stem cells in a first culture medium;

aggregating the stem cells to form a cell pellet; and

culturing the cell pellet in a second culture medium to promote the differentiation of the stem cells of the cell pellet into the insulin producing cells.

3. A method according to claim 2, wherein the stem cells are ones selected from a group consisting of embryonic stem cells, adult stem cells and a combination thereof.

4. A method according to claim 3, wherein the embryonic stem cells are ones selected from a group consisting of embryonic germ cells, transformed embryonic stem cells and induced embryonic stem cells transformed from adult cells and a combination thereof, and the adult stem cells are ones selected from a group consisting of mesenchymal stem cells, hematopoietic stem cells, neural stem cells and a combination thereof.

5. A method according to claim 4, wherein the mesenchymal stem cells are derived from a tissue being one selected from a group consisting of a bone marrow, a cord blood, an adipose tissue and an umbilical cord.

6. A method according to claim 2, wherein the cell pellet includes 2.5×

10⁵ stem cells, and is suspended in the second culture medium.

7. A method according to claim 2, wherein the first culture medium is a complete culture medium including DMEM-low glucose, 10% fetal bovine serum, 100 U/mL penicillin and 10 μ

g/mL streptomycin.

8. A method according to claim 2, wherein the cell pellet is aggregated by centrifuging the stem cells at 200-600 g for 5-15 minutes.

9. A method according to claim 2, further comprising a step of preculturing the cell pellet in the first culture medium before the culturing step.

10. A method according to claim 2, wherein the second culture medium contains one selected from a group consisting of a fibronectin, a laminin and a combination thereof.

11. A method according to claim 2, wherein the culturing step includes further sub-steps of:

culturing the cell pellet for two days at a first stage;

culturing the cell pellet for one day at a second stage;

culturing the cell pellet for four days at a third stage; and

culturing the cell pellet for three days at a fourth stage.

12. A method according to claim 11, wherein:

the second culture medium is a complete culture medium at the first stage;

the second culture medium is a first DMEM/F-12 medium containing insulin-transferrin-selenium-A (ITS-A), 25 mM glucose and 0.45 mM 3-isobutyl-1-methylxanthine (IBMX) at the second stage;

the second culture medium is a second DMEM/F-12 medium containing N2 supplement, B27 supplement, 5.56 mM glucose and 10 mM nicotinamide at the third stage; and

the second culture medium is a third DMEM/F-12 medium containing N2 supplement, B27 supplement, 25 mM glucose and 10 mM nicotinamide at the fourth stage.

13. A method according to claim 2, wherein the insulin producing cells are induced by a glucose to release an insulin, and the glucose has a concentration in a range of 5-25 mM.

14. A method according to claim 13, wherein the release of the insulin is enhanced by an inhibitor of cyclic-AMP phosphodiesterase.

15. A method according to claim 13, wherein the release of the insulin is inhibited by a blocker of Calcium ion channel.

16. An insulin producing cell differentiated from a stem cell, wherein the stem cell is aggregated into a cell pellet to be cultured so as to differentiate into the insulin producing cell.

17. An insulin producing cell according to claim 16, wherein the stem cell is one selected from a group consisting of an embryonic stem cell, an adult stem cell and a combination thereof.

18. An insulin producing cell according to claim 17, wherein the embryonic stem cell is one selected from a group consisting of an embryonic germ cell, a transformed embryonic stem cell and an induced embryonic stem cell transformed from an adult cell and a combination thereof, and the adult stem cell is one selected from a group consisting of a mesenchymal stem cell, a hematopoietic stem cell, a neural stem cell and a combination thereof.

19. An insulin producing cell according to claim 16, wherein the differentiation of the stem cell is promoted by one selected from a group consisting of a fibronectin, a laminin and a combination thereof.

20. An insulin producing cell according to claim 16, wherein the insulin producing cell is induced by a glucose to release an insulin, and the glucose has a concentration in a range of 5-25 mM.