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Method for producing intestinal cells from pluripotent stem cells

A production method and enterocyte technology, which can be applied to non-embryonic pluripotent stem cells, artificially induced pluripotent cells, and gastrointestinal cells, etc., and can solve problems such as small number of cells

Pending Publication Date: 2021-06-11
TOKYO INST OF TECH +2
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Problems solved by technology

However, these hiPSC-derived cells were considered immature due to the low number of cells expressing Lgr5

Method used

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  • Method for producing intestinal cells from pluripotent stem cells
  • Method for producing intestinal cells from pluripotent stem cells
  • Method for producing intestinal cells from pluripotent stem cells

Examples

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Embodiment

[0097] Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to these Examples.

[0098] (A) result

[0099] (1) Collagen Vitrigel supports the differentiation of human iPSCs into intestinal cells characterized by membrane formation and expression of intestinal markers.

[0100] In an attempt to induce the differentiation of human iPS cells into mature intestinal cells, the inventors initially induced embryonic endoderm (DE) cells from human iPSCs on M15 cells. Day 3 (D3) DE cells were dissociated, replaced on CVM inserts, and cultured in M2 medium until day 15. Then, the medium was exchanged into a mature medium (M3, M3-1, or M3-2), and cultured until the 40th day ( figure 1 A. Figure 6 A). Alternatively, day 3 DE cells were repeatedly plated on iMatrix511 precoated plates, cultured in M2 medium until day 10, and subsequently cryopreserved as a day 10 intestinal precursor cell stock. Freeze-thaw the frozen-preserved day 10...

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Abstract

Provided is a method for producing intestinal cells derived from pluripotent stem cells, the produced intestinal cells exhibiting functions which are similar to those of actual intestinal cells. This method for producing intestinal cells uses a culture for differentiation into intestinal cells, which is characterized by containing a GSK3 inhibitor and one type selected from the group consisting of an activating agent for hepatocyte growth factor receptors, adrenal cortex hormone, calcitriol, and dimethyl sulfoxide.

Description

technical field [0001] The present invention relates to a medium for differentiation of enterocytes, a method for producing enterocytes from pluripotent stem cells, and enterocytes produced by the method. Background technique [0002] The intestine is an important tissue for the absorption of nutrients, water, and drugs, and the absorption and metabolism of drugs determines the biological availability of drugs, so it is also important for drug development. For drug development, the human colon cancer cell line Caco-2 is widely used as a model of intestinal epithelium for testing absorption and metabolic functions. However, the activity of metabolic enzymes in Caco-2 cells derived from the large intestine is low, and the activity of its metabolic enzymes is not similar to that of the small intestine (Hubatsch et al.(2007).Nat.Protoc.2,2111-2119; Sun et al.( 2002). Pharm. Res. 19, 4-6). Furthermore, Caco-2 cells have different characteristics among cell lines. Therefore, th...

Claims

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Application Information

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IPC IPC(8): C12N5/071C07K14/78C12N1/00C12N5/074
CPCC12N2501/16C12N5/0679C12N2533/54C12N2501/727C12N2501/12C12N2500/38C12N2501/39C12N2501/999C12N2500/30C12N2502/70
Inventor 粂昭苑白木伸明前田和哉楠原洋之石川晶也渡边辉彦
Owner TOKYO INST OF TECH
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