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Method for producing cartilage cells induced to be differentiated from stem cells

A stem cell differentiation and chondrocyte technology, applied in the field of preparation of chondrocytes induced from stem cell differentiation, can solve problems such as high cost and achieve the effect of high expression level

Pending Publication Date: 2019-10-01
伊普塞尔有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this requires frequent addition of growth factors during differentiation, which has the disadvantage of being expensive

Method used

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  • Method for producing cartilage cells induced to be differentiated from stem cells
  • Method for producing cartilage cells induced to be differentiated from stem cells
  • Method for producing cartilage cells induced to be differentiated from stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0155] Hereinafter, the present invention will be described in detail.

[0156] In the field of regenerative medicine, the technology of inducing the differentiation of chondrocytes by cell therapy technology is widely used for cartilage regeneration, but the therapeutic effect may vary depending on the characteristics of the differentiation ability and tissue formation ability of the transplanted chondrocytes. For this reason, a differentiation technique for inducing chondrocytes derived from reverse-differentiated stem cells is required, and studies are continuing for the purpose of improving differentiation efficiency and tissue-forming ability of differentiated chondrocytes.

[0157] Therefore, the present invention provides a method for preparing chondrocytes induced by stem cell differentiation through the following steps i) to step v):

[0158] Step i), forming (generation) embryoid bodies by culturing reverse differentiated stem cells (induced pluripotent stem cells, i...

Embodiment 1

[0244] Example 1. Generation of human induced pluripotent stem cells using isolated CBMCs

[0245] Reprogramming of CBMCs was facilitated using Sendai virus containing Yamanaka factors. Yamanaka factors are genes that induce pluripotency. After a period of transduction, CBMC-hiPSCs formed colonies similar to embryoid body stem cells ( figure 1 part (a) of this document). CBMC-hiPSCs were purified using a cell line with the same cell morphology. The same CBMC-hiPSCs were used for additional characterization. The identified CBMC-hiPSCs were stained with alkaline phosphate ( figure 1 part (b) of Multidifferentiation markers including OCT4, SOX2, NANOG, LIN28, KLF4, and c-MYC were also assayed ( figure 1 part (c) of Maternal CBMCs were used as a negative control group. The expressions of OCT4, SOX2, NANOG, and LIN28 were increased in CBMC-hiPSCs. However, KLF4 and c-MYC were less expressed in CBMC-hiPSCs than in CBMCs. Typical cell surface markers (SSEA4, OCT4, SOX2, KLF...

Embodiment 2

[0246] Example 2. Chondrocytes differentiated into CBMC-iPSCs

[0247] To confirm the cartilage regeneration ability of CBMC-iPSCs, chondrogenic differentiation was performed by culturing embryoid bodies and inducing growth cells. A simple scheme for the generation process of chondrogenically differentiated cell bodies is shown in figure 2 as shown in part (a). CBMC-iPSCs colonies were prepared for chondrogenic differentiation ( figure 2 part (b) of CBMC-iPSCs expanded and aggregated into embryoid bodies ( figure 2 part (c) of After several days of expansion, the embryoid bodies were transferred to gelatin-coated dishes to induce growth cells ( figure 2 part (d) of this document). Growing cells are expanded and isolated into single cells for chondrocyte differentiation. Utilize 2×10 6iPScs were obtained from many chondrocyte bodies. After 30 days of differentiation, embryoid body growth cells were used to form chondrocyte bodies. The resulting chondrogenic partic...

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Abstract

The present invention relates to a method for inducing differentiation into cartilage cells from dedifferentiated stem cells derived from cord blood mononuclear cells. When a cartilage cell body produced through the method of the present invention is transplanted into an area of cartilage damage in an organism, cartilage regeneration by differentiated cartilage cells may be effectively exhibited.Since more effective cartilage regeneration may be exhibited when the cartilage cell body is transplanted than when cartilage cells induced to be differentiated by adding a recombinant growth factor are transplanted, the cartilage cell body can be usefully used for a tissue engineering treatment method for regenerating cartilage.

Description

technical field [0001] This application claims the priority of Korean Patent Application No. 10-2017-0009015 filed on January 19, 2017 and Korean Patent Application No. 10-2017-0009019 filed on January 19, 2017. The entire content of the above specification is References for this application. [0002] The present invention relates to a method for preparing a composition for differentiating chondrocytes, a composition for differentiating chondrocytes and a composition for differentiating the above-mentioned chondrocytes, or a composition containing chondrogenic differentiation cells for preventing Or a pharmaceutical composition for treating cartilage-related diseases. Background technique [0003] Cartilage is a bone tissue composed of chondrocytes and a cartilage matrix, and refers to a tissue that roughly forms a part of a joint. Most of the cartilage is composed of various collagens, proteoglycans, and flexible fibers rich in chondrocytes and a large amount of extracell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N15/85C07K14/495C07K14/51
CPCC07K14/495C07K14/51C12N5/0655C12N5/0696C12N2501/15C12N2501/155C12N2506/45C12N2506/11C12N2510/00C12N2500/38C12N2501/39C12N2501/105A61K35/32C12N15/85C12N2500/50C12N2501/998C12N2533/54
Inventor 朱址贤南有俊林芮利
Owner 伊普塞尔有限公司
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