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

Active Publication Date: 2014-07-30
JANSSEN BIOTECH INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0017] However, mouse models of embryonic stem cell development may not fully mimic the developmental program in higher mammals such as humans

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
Comparison scheme
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 plates and supplemented with 2% BSA, 100ng / ml activin A, 20ng / ml WNT-3a, 8ng / ml bFGF cultured on 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 co...

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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] This application is a divisional application, the filing date of the original application is July 20, 2010, the application number is 201080033700.0 (PCT / US2010 / 042504), and the title of the invention is "differentiation of human embryonic stem cells". [0002] Cross references to related patent applications [0003] This application claims priority to Application Serial No. 61 / 226,929, filed July 20, 2009. technical field [0004] 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 [0005] 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 ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCA61P3/10C12N5/0678C12N2501/115C12N2501/117C12N2501/15C12N2501/155C12N2501/16C12N2501/385C12N2501/41C12N2501/415C12N2506/02C12N2533/90C12N5/0606C12N5/0676
Inventor J.徐J.延森
Owner JANSSEN BIOTECH INC
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