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Therapeutic agent of peripheral blood flow disorder

A peripheral blood and disorder technology, applied in the field of cell preparations of pluripotent stem cells, can solve the problem of using Muse cells, etc., and achieve the effect of improving or restoring blood flow disorder and excellent safety.

Pending Publication Date: 2021-07-30
KYOTO PREFECTURAL PUBLIC UNIV CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no clear examples of the use of Muse cells in the treatment of peripheral arterial disease and the expected therapeutic effect

Method used

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  • Therapeutic agent of peripheral blood flow disorder
  • Therapeutic agent of peripheral blood flow disorder
  • Therapeutic agent of peripheral blood flow disorder

Examples

Experimental program
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Effect test

preparation example Construction

[0096] (2) Preparation and use of cell preparations

[0097] The cell preparation of the present invention is not particularly limited, and can be obtained by suspending the Muse cells obtained in (1) above or a cell population containing Muse cells in physiological saline or an appropriate buffer (for example, phosphate-buffered saline). In this case, if the number of Muse cells isolated from autologous or allogeneic tissues is small, they can be cultured before cell transplantation until they proliferate to obtain a predetermined number of cells. In addition, as reported (International Publication No. WO2011 / 007900 pamphlet), since Muse cells do not become tumorous, even if the cells recovered from living tissue contain an undifferentiated state, there is little concern about their canceration, So it's safe. In addition, culture of the recovered Muse cells is not particularly limited, and can be performed in an ordinary growth medium (for example, α-minimal essential medium...

Embodiment 1

[0104] Example 1: Preparation of lower limb ischemia model mice

[0105] The experimental protocol using mice in this example complied with the "Kyoto Prefectural University of Medicine Animal Experiments, etc. Relevant Rules", and experimental animals were prepared under the supervision of the Animal Experiment Center of Kyoto Prefectural University of Medicine in accordance with the rules. More specifically, it is prepared by the following steps.

[0106] Female BALB / C nude mice aged 8-10 weeks were anesthetized (induction: 4%, maintenance: 2%) by isoflurane inhalation. The lower limbs were cut open, and the periphery of the left common femoral artery and vein and the left superficial femoral artery and vein were ligated with 5-0 silk thread under a stereomicroscope (KONAN OPERATION MICROSCOPE KOM). The mice thus prepared were used as lower limb ischemia model mice in the following experiments.

Embodiment 2

[0107] Embodiment 2: the preparation of human Muse cell

[0108] Muse cells were obtained according to the method described in International Publication No. WO 2011 / 007900 concerning the isolation and identification of human Muse cells. Commercially available mesenchymal stem cells (MSC, Lonza Corporation) were used as a source of Muse cells. The Muse cells used for transplantation expressed green fluorescent protein (GFP) to confirm that they had attached to the damaged peripheral arterial site, and the lentiviral GFP gene was introduced into the Muse cells in advance to label the cells. GFP-labeled Muse cells were isolated by FACS as double positive cells for GFP and SSEA-3. In addition, the remaining cells from which Muse cells were isolated from MSCs were used as non-Muse cells. In addition, GFP-positive MSCs were also isolated by FACS and used as an MSC panel.

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Abstract

Provided is a cell preparation for treating a peripheral blood flow disorder, the cell preparation including pluripotent stem cells (Muse cells) which are SSEA-3 positive and derived from mesenchymal tissues of a living body or cultured mesenchymal cells. The peripheral blood flow disorder is preferably a peripheral arterial disease, and more preferably, chronic arterial obstruction of the limbs.

Description

technical field [0001] The present invention relates to cell preparations in regenerative medicine. More specifically, the present invention relates to a cell preparation containing pluripotent stem cells effective for angiogenesis therapy of peripheral blood flow disorders such as peripheral arterial disease. Background technique [0002] The proportion of the elderly population aged 65 and over is growing much faster than expected. [0003] On the other hand, with the Westernization of eating habits and changes in lifestyles centered on a mobile society, new concepts of lifestyle-related diseases such as metabolic syndrome have emerged, and the disease structure of society has undergone tremendous changes. In particular, arteriosclerotic diseases and ischemic heart disease are increasing, and at the same time, peripheral arterial disease (PAD: peripheral arterial disease) is also increasing. [0004] PAD mainly refers to chronic arterial occlusive disease of the extremit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P9/00A61P9/08A61K35/545A61P9/10
CPCA61K35/545A61P9/00A61P9/10A61P9/08
Inventor 的场圣明出泽真理
Owner KYOTO PREFECTURAL PUBLIC UNIV CORP
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