Methods and systems for converting precursor cells into intestinal tissues through directed differentiation
US-2017362573-A1 · Dec 21, 2017 · US
Method for producing intestinal cells
US9376665B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9376665-B2 |
| Application number | US-201113882904-A |
| Country | US |
| Kind code | B2 |
| Filing date | Oct 31, 2011 |
| Priority date | Nov 2, 2010 |
| Publication date | Jun 28, 2016 |
| Grant date | Jun 28, 2016 |
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Abstract
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An object of the present invention is to provide a method of producing intestinal cells by use of pluripotent stem cells as a starting material. According to the present invention, provided is a method of producing intestinal cells, comprising the steps of: (A) inducing differentiation of pluripotent stem cells into definitive endoderm cells; and (B) culturing the definitive endoderm cells in the presence of (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO) and N-[(3,5-difluorophenyl)acetyl]-L-Ala-2-phenyl-L-Gly-tert-butyl-OH (DAPT) to thereby induce differentiation of the definitive endoderm cells into intestinal cells.
First claim
Opening claim text (preview).
The invention claimed is: 1. A method of producing intestinal cells expressing Cdx2, comprising the steps of: (a) obtaining mouse or human embryonic stem (ES) cells, (b) culturing the ES cells of step (a) with Activin and/or bFGF to produce definitive endoderm cells, (c) culturing the definitive endoderm cells of step (b) with (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO) and N-[(3,5-difluorophenyl)acetyl]-L-Ala-2-phenyl-L-Gly-tert-butyl-OH (DAPT) and in the presence of M15 cells or MEF cells, to thereby induce differentiation of the definitive endoderm cells into intestinal cells expressing Cdx2. 2. The method according to claim 1 , wherein the definitive endoderm cells are separated from a cell culture obtained in step (b) by flow cytometry using fluorescently-labelled antibodies against E-cadherin (ECD) and CXCR4, and said separated definitive endoderm cells are used in the step (c). 3. The method according to claim 1 , wherein the ES cells are cultured in the presence of feeder cells. 4. The method according to claim 3 , wherein the feeder cells are cells derived from a mesoderm.
Assignees
Inventors
Classifications
connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts" · CPC title
Endothelial cells · CPC title
Notch; Delta; Jagged; Serrate · CPC title
Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10 · CPC title
Small molecules not provided for elsewhere · CPC title
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- What does patent US9376665B2 cover?
- An object of the present invention is to provide a method of producing intestinal cells by use of pluripotent stem cells as a starting material. According to the present invention, provided is a method of producing intestinal cells, comprising the steps of: (A) inducing differentiation of pluripotent stem cells into definitive endoderm cells; and (B) culturing the definitive endoderm cells in t…
- Who is the assignee on this patent?
- Kume Shoen, Ogaki Soichiro, Shiraki Nobuaki, and 3 more
- What technology area does this patent fall under?
- Primary CPC classification C12N5/0679. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jun 28 2016 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).