Differentiation of human embryonic stem cells

A technology of pluripotent stem cells and cells, applied in embryonic cells, artificial cell constructs, pancreatic cells, etc., can solve problems such as not completely simulating the developmental program of higher mammals

Inactive Publication Date: 2012-07-18
JANSSEN BIOTECH INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0016] However, mouse models of embryonic stem cell development may not

Method used

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  • Differentiation of human embryonic stem cells
  • Differentiation of human embryonic stem cells
  • Differentiation of human embryonic stem cells

Examples

Experimental program
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Effect test

example 1

[0144] Human pluripotent stem cells express markers characteristic of the pancreatic endoderm lineage (Table 1 up to PDX1, NKX6.1, but not CDX2 and NGN3) differentiation

[0145] This example demonstrates that activin A can be used in combination with noggin and retinoic acid to promote the upregulation of NKX6.1 expression. Briefly, cells of the human embryonic stem cell line H1 were incubated in MATRIGEL TM (1:30 dilution) coated plate and supplemented with 2% BSA, 100ng / ml activin A, 20ng / ml WNT-3a, 8ng / ml bFGF cultured on the RPMI medium for one day, and then supplemented with 2%BSA , 100ng / ml activin A, 8ng / ml bFGF in RPMI medium for two days (phase 1), then

[0146]a. Treated with DMEM / F12+2%BSA+50ng / ml FGF7+0.25μM cyclopamine-KAAD for three days (stage 2), then

[0147] b. Use DMEM-high glucose+1% B27+50ng / ml FGF7+0.25μM cyclopamine-KAAD+2μM retinoic acid (RA)+100ng / ml Noggin+20ng / ml Activin A or 50ng / ml Activin A treatment for four days (phase 3).

[0148] As a...

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Abstract

The present invention provides methods to promote the differentiation of pluripotent stem cells into cells expressing markers characteristic of the pancreatic endocrine lineage that co-express PDX1, NKX6.1, but do not express CDX2 and NGN3.

Description

[0001] Cross references to related patent applications [0002] This application claims priority to Application Serial No. 61 / 226,929, filed July 20, 2009. technical field [0003] The present invention provides methods to promote the differentiation of pluripotent stem cells into cells expressing markers characteristic of the pancreatic endocrine lineage, co-expressing PDX1, NKX6.1, but not CDX2 and NGN3. Background technique [0004] Advances in cell replacement therapy for type I diabetes and the scarcity of transplantable islets have focused attention on developing a source of insulin-producing cells or beta cells suitable for engraftment. One approach is to generate functional beta cells from pluripotent stem cells, such as embryonic stem cells. [0005] In vertebrate embryonic development, pluripotent stem cells can give rise to a cell population comprising the three germ layers (ectoderm, mesoderm and endoderm) in a process called gastrulation. Tissues such as the t...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0735C12N5/02
CPCC12N2501/15C12N2501/155C12N2501/117C12N2501/115C12N2506/02C12N2533/90C12N5/0678C12N2501/415C12N2501/16C12N2501/385C12N2501/41A61P3/10C12N5/0606C12N5/0676
Inventor J·徐J·延森
Owner JANSSEN BIOTECH INC
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