| US 7,413,901 B2 | ||
| Method of forming vertebrate pancreas in vitro | ||
| Makoto Asashima, Tokyo (Japan); and Naomi Moriya, Mitaka (Japan) | ||
| Assigned to Japan Science and Technology Agency, Kawaguchi-shi (Japan) | ||
| Appl. No. 10/258,712 PCT Filed Mar. 13, 2001, PCT No. PCT/JP01/01963 § 371(c)(1), (2), (4) Date Oct. 25, 2002, PCT Pub. No. WO01/83712, PCT Pub. Date Nov. 08, 2001. |
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| Claims priority of application No. 2000-128336 (JP), filed on Apr. 27, 2000. | ||
| Prior Publication US 2003/0186434 A1, Oct. 02, 2003 | ||
| Int. Cl. C12N 5/00 (2006.01); C12N 5/02 (2006.01); A61F 2/02 (2006.01) | ||
| U.S. Cl. 435—377 [435/325; 435/375; 424/93.1; 623/23.72] | 6 Claims |
| 1. A method of forming an explant that expresses insulin in vitro, comprising:
(a) isolating a Spemann-Mangold organizer region from a vertebrate gastrula;
(b) culturing said Spemann-Mangold organizer region for a predetermined period of time, wherein the period is determined by
a comparison of pancreatic differentiation levels of said Spemann-Mangold organizer region which is cultured for different
periods of time and treated with retinoic acid;
(c) treating said Spemann-Mangold organizer region with retinoic acid of a predetermined concentration for a predetermined
period of time, wherein the concentration is determined by a comparison of the pancreatic differentiation levels of said Spemann-Mangold
organizer region which are treated with retinoic acid of different concentrations, and the period is determined by a comparison
of pancreatic differentiation levels of said Spemann-Mangold organizer region which is treated with retinoic acid; and
(d) culturing said Spemann-Mangold organizer region to produce an explant that expresses insulin.
|