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Undifferentiated stem cell-removing agent, and method for removing undifferentiated stem cells

A technology for differentiated cells and stem cells, applied in the field of undifferentiated stem cell remover, to achieve efficient removal effect

Pending Publication Date: 2019-07-23
KEIO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in vitro, it is difficult to induce differentiation of all stem cells into the target lineage

Method used

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  • Undifferentiated stem cell-removing agent, and method for removing undifferentiated stem cells
  • Undifferentiated stem cell-removing agent, and method for removing undifferentiated stem cells
  • Undifferentiated stem cell-removing agent, and method for removing undifferentiated stem cells

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0167] "Methods for preparing cells for transplantation"

[0168] In one embodiment, the present invention provides a method for preparing cells for transplantation, comprising the following steps (i) and (ii):

[0169] Step (i): inducing desired differentiated cells from undifferentiated stem cells; and

[0170] Step (ii): culturing the cell mixture obtained in step (i) in the presence of an undifferentiated stem cell removing agent.

[0171] The undifferentiated stem cell-removing agent used in the method for preparing cells for transplantation of this embodiment is the undifferentiated stem cell-removing agent of the above-mentioned embodiment.

[0172] Step (i) in the method of this embodiment refers to the step of inducing desired differentiated cells from undifferentiated stem cells. The undifferentiated stem cells in step (i) are not particularly limited, preferably iPS cells or ES cells, more preferably iPS cells.

[0173] In step (i), the type of differentiated cel...

experiment example 9

[0275] [Experimental Example 9] Cultivation of an undifferentiated stem cell line in a medium containing a fatty acid synthesis inhibitor and palmitic acid

[0276] Orlistat (manufactured by Sigma, O4139) was added to StemFit medium (manufactured by AJINOMOTO) or mTeSR1 (manufactured by Stem Cell Technologies) so that each final concentration became as follows Figure 13 Concentrations indicated (2 μM, 6 μM and 8 μM). Thus, a culture medium according to one embodiment of the present invention was obtained. By adding palmitic acid (50 μM final concentration), carnitine (0.5 mM final concentration) and BSA (final concentration 8.3 μM) was used to prepare the culture medium. By adding only carnitine (final concentration 0.5 mM) and BSA (final concentration 8.3 μM), the culture medium was prepared. The human iPS cell line (253G4) was cultured in each of the above media for 72 hours.

[0277] After culturing for 72 hours, live cells were detected with LIVE / DEAD Assay (L3224, m...

experiment example 11

[0285] [Experimental Example 11] Acidic culture of FASN KD undifferentiated stem cell line in medium containing palmitic acid

[0286] A medium obtained by adding palmitic acid (50 μM final concentration), carnitine (0.5 mM final concentration) and BSA (8.3 μM final concentration) to StemFit medium (manufactured by AJINOMOTO) or mTeSR1 (manufactured by Stem Cell Technologies) (PA-BSA medium) and by adding only carnitine (final concentration 0.5 mM) and BSA (final concentration 8.3 μM) to StemFit medium (manufactured by AJINOMOTO) or mTeSR1 (manufactured by Stem Cell Technologies) The obtained medium (BSA medium) was prepared. The FASN KD undifferentiated cell line and the N / C undifferentiated cell line prepared in Experimental Example 11 were cultured in each of the above-mentioned media for 72 hours to confirm cell proliferation. The result is as Figure 17 shown.

[0287] based on Figure 17 Results shown in , FASN KD undifferentiated stem cell line cultured in PA-BSA medi...

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Abstract

An undifferentiated stem cell-removing agent which comprises at least one member selected from the group consisting of a fatty acid synthesis inhibitor, a fatty acid utilization inhibitor and a cholesterol synthesis inhibitor. A method for removing undifferentiated stem cells, the method comprising culturing a cell mixture containing undifferentiated stem cells and differentiated cells in the presence of the aforesaid undifferentiated stem cell-removing agent. A method for producing cells for transplantation, the method comprising: (i) a step for inducing desired differentiated cells from undifferentiated stem cells; and (ii) a step for culturing the cell mixture obtained in step (i) in the presence of the aforesaid undifferentiated stem cell-removing agent.

Description

technical field [0001] The present invention relates to an agent for removing undifferentiated stem cells and a method for removing undifferentiated stem cells. The present invention further relates to a culture medium containing an undifferentiated stem cell removing agent, a method for preparing cells for transplantation, and a pharmaceutical composition. [0002] Priority is claimed from Japanese Patent Application No. 2016-203839 filed on October 17, 2016, the contents of which are incorporated herein by reference. Background technique [0003] In recent years, research on pluripotent stem cells such as embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells) has progressed. Because these cells are pluripotent, cells differentiated into desired lineages can be produced by differentiation and induction, and the resulting cells can be used in medical transplantation. [0004] Differentiation and induction of embryonic stem cells and induced pluripot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K31/198A61K31/336A61K31/365A61K31/4458A61K31/7004A61K35/12A61K35/34A61K45/00A61K45/06A61P43/00C12N5/10C12N9/99
CPCC12N9/99A61K35/34C12Y203/01021C12Y203/01085C12Y203/03008C12Y401/01009C12Y604/01002C12N2506/02C12N2506/45C12N5/0657C12N2501/70C12N2501/73A61L27/3604A61P9/00C12N5/0081A61L2430/20C12N2501/72C12N2501/71A61K31/198A61K31/336A61K31/365C12N5/00C12N5/0018C12N5/0696A61K31/7004A61K35/12C12N5/10A61K31/4458A61P43/00A61K45/00A61K45/06
Inventor 远山周吾福田恵一藤田淳田野崎翔
Owner KEIO UNIV