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Differentiation-inducing culture medium additive and use thereof

a technology additive, which is applied in the field of differentiation-inducing culture medium additive, can solve the problems of high cost of serum, difficult to achieve the effect of low cos

Inactive Publication Date: 2011-09-01
HIROSHIMA UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0038]As described above, a differentiation-inducing culture medium additive of the present invention for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition, at least contains: at least one growth factor selected from the group consisting of EGF, FGF, and PDGF; dexamethasone; and β-glycerophosphate. Accordingly, by adding the differentiation-inducing culture medium additive of the present invention to a basal medium, it is possible to produce an effect that bone differentiation of a target cell can be efficiently induced under a serum-free condition and an effect that use of serum, which may include a pathogen and may cause variation in differentiation, can be avoided. Further, recombinant human insulin, transferrin, selenate, and ascorbic acid, which have been conventionally considered as being essential for bone differentiation induction, need not necessarily to be added. This makes it possible to provide a differentiation-inducing culture medium additive at lower cost.

Problems solved by technology

However, a serum is very expensive, and components thereof differ for each lot because the serum is a natural product.
This requires a complicated process.
Furthermore, there is a risk that cultured cells may be infected with an unknown pathogen (such as a virus or pathogenic prion) that is included in the serum.
This may cause bone differentiation of the mesenchymal stem cell to be carried out with lower efficiency.

Method used

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  • Differentiation-inducing culture medium additive and use thereof
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  • Differentiation-inducing culture medium additive and use thereof

Examples

Experimental program
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example 1

Study on Differentiation of Mesenchymal Stem Cells into Osteoblast Cells (1)

[0130]Three human ilium-marrow-derived mesenchymal stem cell lines (hereinafter referred to as a cell line A, a cell line B, and a cell line C) (purchased from Bio-Whittaker Inc. (Walkersville, Md.)) were used as mesenchymal stem cells. Note that the cell line A is a fifth subculture of human ilium-marrow-derived mesenchymal stem cells and each of the cell lines B and C is a fourth subculture of human ilium-marrow-derived mesenchymal stem cells. A DMEM culture medium containing 10% FBS was used to culture the mesenchymal stem cells. The mesenchymal stem cells were seeded on a 6-well microplate containing the DMEM culture medium at a density of 5000 cells / cm2. Culture media were replaced every two or three days during the cell culture.

[0131]Note that the DMEM culture medium containing 10% FBS was prepared by adding fetal bovine serum (FBS) in a final concentration of 10% by weight to DMEM (produced by Sigma A...

example 2

Study on Differentiation of Mesenchymal Stem Cells into Osteoblast Cells (2)

[0146]Five human ilium-marrow-derived mesenchymal stem cell lines (hereinafter referred to as a cell line F, a cell line G, a cell line H, a cell line I, and a cell line J) (purchased from Bio-Whittaker Inc. (Walkersville, Md.)) were used as mesenchymal stem cells. Note that each of the cell lines F, H, and I is a fifth subculture of human ilium-marrow-derived mesenchymal stem cells and each of the cell lines G and J is a fourth subculture of human ilium-marrow-derived mesenchymal stem cells. A DMEM culture medium containing 10% FBS was used to proliferate and culture the mesenchymal stem cells. The mesenchymal stem cells were seeded on a 6-well microplate containing the DMEM culture medium containing 10% FBS at a density of 5000 cells / cm2. Culture media were replaced every two or three days during the cell culture.

[0147]Before becoming confluent, the mesenchymal stem cells were collected from the 6-well mic...

example 3

Study on Differentiation of Mesenchymal Stem Cells into Osteoblast Cells (3)

[0161]Three human ilium-marrow-derived mesenchymal stem cell lines (hereinafter referred to as a cell line K, a cell line L, and a cell line M) (purchased from Bio-Whittaker Inc. (Walkersville, Md.)) were used as mesenchymal stem cells. Note that the cell line K is an eighth subculture of human ilium-marrow-derived mesenchymal stem cells, the cell line L is a seventh subculture of human ilium-marrow-derived mesenchymal stem cells and the cell line M is a tenth subculture of human ilium-marrow-derived mesenchymal stem cells.

[0162](1. Induction of Bone Differentiation of Mesenchymal Stem Cells Proliferated and Cultured by Use of Serum-Free STK2 Culture Medium)

[0163]The mesenchymal stem cells were seeded on a 6-well microplate containing a serum-free STK2 culture medium at a density of 5000 cells / cm2. Culture media were replaced every two or three days during the cell culture.

[0164]The mesenchymal stem cells we...

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Abstract

Provided is a differentiation-inducing culture medium additive for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition, and a use of the differentiation-inducing culture medium additive. The differentiation-inducing culture medium additive of the present invention for inducing differentiation of a stem cell under a serum-free condition at least contains at least one growth factor selected from the group consisting of EGF, FGF, and PDGF; dexamethasone; and β-glycerophosphate. The differentiation-inducing culture medium additive of the present invention does not require ascorbic acid 2-phosphate and ITS, which are normally essential for bone differentiation. Further, bone differentiation can be promoted by adding phospholipid.

Description

TECHNICAL FIELD[0001]The present invention relates to a differentiation-inducing culture medium additive and a use thereof. More specifically, the present invention relates to a differentiation-inducing culture medium additive for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition, and to a use of the differentiation-inducing culture medium additive.BACKGROUND ART[0002]A stem cell is a cell having a self-reproducing potential and a differentiation potential. For example, an embryonic stem cell (an ES cell), an induced pluripotent stem cell (an iPS cell), and a somatic stem cell are known as stem cells. Examples of the somatic stem cell include: a hematopoietic stem cell, a neural stem cell, and a mesenchymal stem cell. Each of the ES cell and the iPS cell has a pluriopotency to differentiate into any tissue. ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2500/25C12N2500/99C12N2501/11C12N2501/39C12N2501/12C12N2501/135C12N2501/15C12N2501/115C12N2500/42C12N2500/90C12N2501/40C12N2506/1353
Inventor KATO, YUKIOSHAO, JIN CHANGTSUJI, KOICHIRO
Owner HIROSHIMA UNIVERSITY
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