Method for inducing tailored pluripotent stem cells using extract of plant stem cells or plant dedifferentiated stem cells, and pluripotent stem cells produced by means of the method

Patent metadata

Field	Value
Publication number	US-9976118-B2
Application number	US-201414762773-A
Country	US
Kind code	B2
Filing date	Jan 28, 2014
Priority date	Jan 30, 2013
Publication date	May 22, 2018
Grant date	May 22, 2018

Abstract

A method for inducing tailored pluripotent stem cells by reprogramming differentiated cells of an adult by using an extract of plant stem cells and plant dedifferentiated stem cells (callus); pluripotent stem cells produced by means of the method; and a cell therapy agent comprising the pluripotent stem cells are disclosed. It is possible to overcome ethical problems since eggs are not used in the making of pluripotent stem cells having abilities like embryonic stem cells. It is possible to produce pluripotent stem cells of ensured safety because a plant stem cell extract which above all has been verified to be harmless to the body is used. It is possible to develop an immunocompatible cell therapy agent tailored to different individuals.

First claim

The invention claimed is:

1. A method of producing induced stem cells, comprising:
pre-treating human dermal fibroblasts with a membrane permeability-promoting material;
contacting a Sequoia callus extract with the pre-treated human dermal fibroblasts such that at least part of the Sequoia callus extract is permeated into the pre-treated human dermal fibroblasts, which provides modified human cells containing at least part of the Sequoia callus extract;
culturing, in a culture medium, the modified human cells containing at least part of the Sequoia callus extract, which produces induced stem cells expressing Nanog and Oct-3/4; and
subsequently collecting at least part of the induced stem cells expressing Nanog and Oct-3/4.

2. The method of claim 1 , wherein the Sequoia callus extract comprises proteins, lipids, sugars and acids of callus of Sequoia.

3. The method of claim 1 , wherein the method further comprises mixing the Sequoia callus extract with the culture medium before contacting the Sequoia callus extract with the human dermal fibroblasts.

4. The method of claim 1 , wherein the Sequoia callus extract contacts the pre-treated human dermal fibroblasts in the form of suspension in water and has a concentration of 100 ug/ml to 1 mg/ml.

5. The method of claim 4 , wherein the concentration is 500 ug/ml.

6. The method of claim 1 , wherein the Sequoia callus extract is in powder.

7. The method of claim 6 , wherein the powder of the Sequoia callus extract is dissolved in water before contacting the pre-treated human dermal fibroblasts.

8. The method of claim 1 , wherein the membrane permeability-promoting material includes streptolysin O.

9. The method of claim 1 , wherein the membrane permeability-promoting material includes digitonin.

10. The method of claim 1 , wherein culturing comprises transferring the modified human cells containing at least part of the Sequoia callus extract onto a surface of feeder cells.

11. The method of claim 10 , wherein the feeder cells comprise STO cells.

12. The method of claim 10 , wherein the culture medium comprises a Knockout serum replacement.

13. The method of claim 10 , wherein subsequent to transferring, the modified human cells containing at least part of the Sequoia callus extract are cultured on the surface of the feeder cells, wherein the culture medium comprises a Knockout serum replacement, wherein culturing in the culture medium is performed subsequent to culturing on the surface of the feeder cells.

14. The method of claim 1 , wherein the culture medium comprises a Knockout serum replacement.

Assignees

• Snu R&Db Foundation

Inventors

• Kwon Yoo-Wook
• Park Young-Bae
• Kim Hyo-Soo
• Paek Jae-Seung

Classifications

• A61K2800/86

  Products or compounds obtained by genetic engineering · CPC title

• C12N5/04 Primary

  Plant cells or tissues {(culture media C12N5/0025)} · CPC title

• A61K2800/80

  Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof · CPC title

• A61K8/97

  from algae, fungi, lichens or plants; from derivatives thereof · CPC title

• C12N5/0696

  Artificially induced pluripotent stem cells, e.g. iPS · CPC title