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Pharmaceutical agent for promoting functional regeneration of damaged tissue

A technology of tissue regeneration and accelerators, applied in the direction of microorganisms, drug combinations, tissue culture, etc., can solve the problems of poor survival prognosis and insufficient effect of malignant glioma

Inactive Publication Date: 2011-05-25
GENOMIX CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the survival prognosis of malignant glioma is poor, and neither chemotherapy nor radiation therapy nor immunology and gene therapy, which have been widely studied in recent years, have shown sufficient effects

Method used

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  • Pharmaceutical agent for promoting functional regeneration of damaged tissue
  • Pharmaceutical agent for promoting functional regeneration of damaged tissue
  • Pharmaceutical agent for promoting functional regeneration of damaged tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0168] Objective: To evaluate the involvement of bone marrow-derived cells in the functional regeneration of transplanted skin tissue in vivo

[0169] Method: For the purpose mentioned above, research was carried out by the following method.

[0170] 1) Using the in vivo skin transplantation system implanted into GFP bone marrow transplanted mice, the degree of participation of bone marrow-derived cells in the functional regeneration of the grafted skin was studied. Specifically, C57BL / 6 male mice (6-8 weeks old) were irradiated with a lethal dose of radiation (10Gy), and then transplanted bone marrow cells (5×10 6 each / 0.1ml physiological phosphate buffer solution pH7.4)( figure 1 ).

[0171] 2) After transplanted bone marrow cells were implanted (6 weeks), newborn mouse skin (female) was transplanted on the back skin of the obtained GFP bone marrow transplanted mouse.

[0172] 3) After the grafted skin is implanted and the skin tissue is fully regenerated (4 weeks), the c...

Embodiment 2

[0178] Objective: To identify bone marrow-derived tissue stem cell-inducing factors in skin tissue extracts

[0179]METHODS: In order to identify the bone marrow mesenchymal stem cell mobilization factors presumed to be released from the resected skin in a state of hypoperfusion, the following study was carried out.

[0180] 1) To obtain mouse bone marrow-derived mesenchymal stem cells, mouse bone marrow cells were collected from the femur or lower leg bone of a C57BL / 6 mouse, and D-MEM (manufactured by Nacalai) containing 10% fetal bovine serum was used as The cell culture medium was added to the cell culture dish, and cultured at 37° C. and a carbon dioxide gas concentration of 5%. When the cells proliferate to an area of ​​70-100% relative to the bottom area of ​​the culture dish, the cells are detached from the culture dish with 0.25% trypsin 1mM EDTA (manufactured by Nacalai), and then subcultured under the same conditions. to cultivate. The subculture operation was rep...

Embodiment 3

[0192] Objective: To confirm the therapeutic effect of S100A8 on skin ulcer in normal mice and diabetic mice

[0193] Method: The recombinant S100A8 protein was given to the mouse skin ulcer model to study the effect of ulcer treatment. As the test mice, C57 / B16 mice transplanted with GFP-expressing bone marrow cells or diabetic model mice BKS.Cg-m+ / +Leprdb / J (db mice) were used. Skin ulcers with a diameter of 6 mm were made on the mouse skin. The skin around the skin defect portion of the mouse-made skin ulcer shrank rapidly. In this experiment, in order to create a model in which the defective skin does not shrink but is treated by covering it with regenerated skin, a silicone rubber disc with an outer diameter of 10 mm, an inner diameter of 6 mm, and a thickness of 0.5 mm was used for skin surgery. Adhesive (Aron alpha A) and nylon thread are fixed to the skin around the ulcer. Then, the recombinant S100A8 protein was directly administered to the ulcer surface at 1.5 μg / ...

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Abstract

Studies have been made on a possibility that a bone-marrow-derived cell is recruited from an extracutaneous tissue into a skin graft during the process of adhesion of the skin graft to a biological tissue and therefore contributes to the regeneration of a skin tissue. As a result, the following facts 1) to 4) are found: 1) a large quantity of bone-marrow-derived cells are recruited into a skin graft; 2) the recruited bone-marrow-derived cells are differentiated into all of dermal fibroblasts, adipocytes, muscle cells, vascular endothelial cells and epidermal keratinocytes in the skin graft, and bone-marrow-derived mesenchymal stem cells are contained in the recruited bone-marrow-derived cells; 3) the substances which recruit the bone-marrow-derived mesenchymal stem cells from the peripheral blood into the skin graft are S100A8 and S100A9 which are released from a dead tissue in the skin graft; and 4) purified S100A8 and S100A9 can promote the migration of a mesenchymal stem cell which has been separated from a bone marrow and cultured.

Description

technical field [0001] The invention relates to functional regeneration-promoting drugs for damaged tissues. Background technique [0002] In recent years, it has become clear that various stem cells are involved in the repair process of damaged tissue, and the development of new regenerative medicine technology that induces functional tissue regeneration by mobilizing a large number of stem cells to the damaged site is also proceeding. In order to make this new regenerative medicine technology a reality, 1) there are abundant stem cells in the body that can mobilize to the damaged site, and 2) the factors that mobilize the stem cells to the damaged site must be isolated and identified. [0003] Stem cells that can be mobilized to the injury site include tissue stem cells present in the injury site or nearby tissues and bone marrow-derived stem cells present in peripheral blood. In recent years, it has been continuously reported that bone marrow-derived cells participate in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/00A61K31/7088A61K35/76A61K48/00A61P17/02A61P43/00C07K14/47C12N5/00C12N15/09
CPCC12N15/8509A01K2267/0393C07K14/47A01K2227/105A61K38/1709A61K31/7088A61P17/02A61P43/00
Inventor 玉井克人山崎尊彦知野刚直金田安史
Owner GENOMIX CO LTD
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