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Treatment agent for epidermolysis bullosa

A technology for bullous and lysis, which is applied in the field of cell preparations, can solve problems such as undisclosed treatment effects, and achieve the effect of improving or restoring skin symptoms and having good safety.

Pending Publication Date: 2020-02-21
HOKKAIDO UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no disclosed example of using Muse cells for the treatment of epidermolysis bullosa to obtain the desired therapeutic effect

Method used

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  • Treatment agent for epidermolysis bullosa
  • Treatment agent for epidermolysis bullosa
  • Treatment agent for epidermolysis bullosa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Example 1: Evaluation of full-thickness trauma model mice

[0125] Adult C57BL / 6 mice were used as a model by performing full-thickness wounds on the back. Within 30 minutes from the implementation of the full-thickness wound, the Muse cells prepared above (3×10 5 / mouse or 3 x 10 4 / mouse), MSC (3×10 5 / mouse) or HBSS 200μl for tail vein injection to test the therapeutic effect.

[0126] In addition, the epithelialization rate was calculated as follows.

[0127] Use a ruler and a digital camera to take pictures of the back skin at the time of wound creation and the 3rd, 6th, 9th, and 11th day after wounding, and use Image J software (version 1.50i) to calculate the respective skin ulcer areas (mm 2 ). Taking the area at the time of wound creation as a reference value, the percentage (%) of area reduction rate was calculated.

[0128] [Formula 1]

[0129]

[0130] The result is as figure 1 with figure 2 shown. In either group, wounds healed over time, but ...

Embodiment 2

[0132] Example 2. Evaluation of COL17 knockout epidermolysis bullosa model mice

[0133] In COL17 knockout mice (reference Nat Med. 2007 Mar; 13(3): 378-83.) (3 to 4 weeks old), blisters are formed by rubbing the epidermis, and injected through the tail vein within 30 minutes from the formation of blisters Muse cells (3×10 5 / mouse). As a result of observing the state of the skin after one month, as Figure 4 As shown, in the control mice administered with HBSS, the state of the coat was poor, and the formation of wounds and mucosal erosions was widely confirmed, but in the mice administered with Muse cells, the state of the coat and the formation of wounds were relatively poor. light. In addition, skin tissue one month after the application was isolated, RNA was extracted therefrom, and expression of human-derived COL7 gene and COL17 gene was examined by RT-PCR. The result is shown in 5. As a result, the presence of human COL7 and human COL17 was confirmed in mice admini...

Embodiment 3

[0134] Example 3. Induction of differentiation of Muse cells into keratinocytes

[0135] Differentiation into keratinocytes was induced by culturing Muse cells according to the following procedure.

[0136] Inoculation of Muse cells on day 0

[0137] On the first day, culture with DMEM low glucose medium + 10% FBS + KGF (10ng / ml) + EGF (20-30ng / ml) for 3 days

[0138] On day 4, culture with DMEM low glucose medium + 10% FBS + KGF (10ng / ml) + EGF (20-30ng / ml) + HGF (10ng / ml) + IGF2 (60ng / ml), every other day Replace the medium once and culture for 8-14 days

[0139] The result is as Figure 7-9 shown. Such as Figure 7 As shown, cells at day 8 of differentiation exhibited a keratinocyte-like morphology. In addition, if Figure 8 with Figure 9 As shown, differentiation-induced cells showed expression of keratinocyte markers at both protein and mRNA levels.

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Abstract

Provided is a cell preparation for treating epidermolysis bullosa, containing SSEA-3 positive pluripotent stem cells (muse cells) derived from a mesenchymal tissue of a live body or cultured mesenchymal cells. The epidermolysis bullosa is preferably epidermolysis bullosa simplex, junctional epidermolysis bullosa, or dystrophic epidermolysis bullosa.

Description

technical field [0001] The invention relates to a cell preparation for regenerative medicine. More specifically, it relates to a cell preparation containing pluripotent stem cells effective for the treatment of epidermolysis bullosa and a cell preparation containing pluripotent stem cells effective for the treatment of epidermolysis bullosa and other skin diseases induced by differentiation. The resulting cell preparation of skin cells. Background technique [0002] Epidermolysis bullosa (Epidermolysis bullosa, EB) is the destruction of the adhesion function between the epidermis and dermis due to the genetic abnormality of the adhesion structure control protein in the basement membrane area of ​​the skin. Hereditary vesicular skin disease that peels off at the basement membrane level to form scalded blisters and ulcers all over the body (Table 1). Epidermolysis bullosa can be roughly divided into three types: simple type, junctional type, and dystrophic type according to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/545A61K35/28A61K35/33A61K35/36A61P17/00A61P17/02C12N5/071C12N5/0775
CPCA61K35/28A61K35/33A61K35/36A61K35/545A61P17/02C12N5/0629C12N5/0656C12N2506/03A61P17/00C12N5/0625
Inventor 清水宏藤田靖幸桝富直哉
Owner HOKKAIDO UNIVERSITY
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