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Encapsulated intestinal midgut endoderm cells

a technology of endoderm cells and endoderm cells, which is applied in the field of cell-based therapy for conditions, can solve the problems of inefficient process of differentiating enteroendocrine cells from these hesc-derived mid-/hindgut spheroids, and no incretin-based cell therapy option,

Inactive Publication Date: 2019-12-05
JANSSEN BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention provides a method for generating intestinal midgut endoderm cells from human pluripotent stem cells. The method involves inducing differentiation of the stem cells into definitive endoderm cells, then culturing them in a specific media to induce differentiation into primitive gut tube cells, and finally culturing them in a different media to induce differentiation into intestinal midgut endoderm cells. These cells express specific markers and transcription factors characteristic of intestinal midgut endoderm cells, and can be used for treating diabetes. The invention also provides a population of intestinal midgut endoderm cells that form a monolayer in culture.

Problems solved by technology

However, there is currently no incretin-based cell therapy option that would encompass an endogenous and cellular barometer for improved and efficient GLP1-based diabetes treatment.
Furthermore, current incretin-based therapies are not regulated by circulating blood glucose levels and thus provide non-physiologically regulated GLP production.
The process of differentiating enteroendocrine cells from these hESC-derived mid- / hindgut spheroids is very inefficient, requiring a long time period, and is directed non-discriminately towards the generation of all intestinal cell types of the intervillus and villus regions.

Method used

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  • Encapsulated intestinal midgut endoderm cells
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  • Encapsulated intestinal midgut endoderm cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

Method of Producing an Intestinal Midgut Endoderm Cell Population with CDX2 and FOXA2 Co-Presence / Co-Expression

[0110]The following example describes a directed-based method to generate intestinal midgut endoderm cells from human embryonic stem cell (“hESC”). “Intestinal midgut endoderm” refers to a corresponding in vivo or in situ cell type which is CDX2-positive and FOXA2-positive endoderm cells present at about embryonic day 8.5 (“E8.5”) during mouse development, or at about the 3-4 week time point during human embryonic development.

Materials and Methods

[0111]Cell culture: Cells of the human embryonic stem cell line H1 (“H1-hESC”) (WA01 cells, WiCell Research Institute, Madison, Wis.) cultured with EZ8 media (Cat #A1516901 Gibco, Thermo Fisher Scientific) at passage 28 were seeded as single cells at 0.094×106 cells / cm2 on MATRIGEL™, at a 1:30 dilution, (Corning Incorporated, Corning, N.Y., Catalog #356231) coated dishes in a media of Dulbecco's Modified Eagle's Medium Nutrient mix...

example 2

Intestinal Culturing Starting from the Definitive Endoderm, and Using FGF4 and WNT-Agonists, Generates an Endoderm-Mesenchyme Mixture of CDX2+ Mid- / Hindgut Cells

[0131]This example demonstrates the endoderm-mesenchyme-mixed quality of the CDX2+ mid- / hindgut cells generated from intestinal culturing beginning at the definitive endoderm stage using FGF4 and WNT-agonists (Spence et al., Nature, 2011; 470:105-109; Watson et al. Nature Med, 2014; 11:1310-1314). To examine the induction to midgut / hindgut endoderm cells described in Spence et al., infra, hESCs were differentiated using the protocol below. Note that the differentiation conditions outlined in this Example differ from Example 1 by the following: (i) intestinal condition starting point begins at the definitive endoderm stage; (ii) different growth factors and small molecules are used than RA and BMP4 or BMP2; and (iii) acidic culture conditions are not used.

Materials and Methods

[0132]Cell culture: H1-hESC cells were cultured an...

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Abstract

Cell populations of intestinal midgut endoderm cells and methods of generating the cells expressing markers characteristic of intestinal endoderm lineage are disclosed. Methods of treating conditions such as diabetes are also disclosed.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. application Ser. No. 15 / 478,881, filed Apr. 4, 2017, which claims the benefit of U.S. Provisional Application No. 62 / 322,636, filed Apr. 14, 2016. The above-listed applications are herein incorporated by reference in their entirety.FIELD OF INVENTION[0002]The invention relates to the field of cell-based therapy for conditions such as diabetes. In particular, the invention relates to cell differentiation, including directing differentiation of human pluripotent stem cells to generate a population of intestinal midgut endoderm cells. The invention provides cells or a cell population and methods of producing the cells that express markers characteristic of intestinal midgut endoderm.BACKGROUND[0003]Advances in the knowledge of incretin hormone mechanism of action coupled with advancements in the understanding of intestinal differentiation, both at the stem cell and endocrine cell stages, have led to...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/38A61K35/545C12N5/071
CPCC12N2500/38C12N2501/155C12N2501/119C12N5/0679A61K35/38A61K35/545C12N2501/19C12N2501/727C12N5/0603A61P3/10C12N2506/45C12N2501/16C12N2501/117C12N2500/30C12N2501/30C12N2501/60C12N2501/602C12N2500/60C12N2501/385C12N2501/415C12N2506/02C12N2501/72C12N2501/10C12N5/00
Inventor RIECK, SEBASTIANREZANIA, ALIREZA
Owner JANSSEN BIOTECH INC
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