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Composition for allotransplantation cell therapy, said composition containing SSEA-3 positive pluripotent stem cell capable of being isolated from body tissue

A technology of pluripotent stem cells and cell therapy, applied in the field of cell therapy compositions for allogeneic transplantation, which can solve the problems of unclear cells and low differentiation efficiency

Inactive Publication Date: 2013-12-11
克里奥公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the bone marrow mesenchymal cell part is a cell population containing multiple cell types, and the differentiation efficiency is not high when induction is applied.
It can be speculated that some cells in MSC are responsible for differentiation, but the above-mentioned cells themselves have not been clearly identified, and this has been a subject of long-term discussion

Method used

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  • Composition for allotransplantation cell therapy, said composition containing SSEA-3 positive pluripotent stem cell capable of being isolated from body tissue
  • Composition for allotransplantation cell therapy, said composition containing SSEA-3 positive pluripotent stem cell capable of being isolated from body tissue
  • Composition for allotransplantation cell therapy, said composition containing SSEA-3 positive pluripotent stem cell capable of being isolated from body tissue

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0196] Example 1 Expression of HLA antigens in Muse cells

[0197] The expression of HLA class I and HLA class II antigens in SSEA-3 positive cells derived from human bone marrow mesenchymal cells was confirmed by flow cytometry. figure 1 Indicates the result. Such as figure 1 As shown, in human bone marrow mesenchymal cells, HLA1 is positive, but HLA2 is negative.

[0198] The expression of HLA class I and HLA class II antigens in human fibroblast-derived SSEA-3 positive cells was also confirmed. figure 2 Indicates the result. Such as figure 2 As shown, in human fibroblasts, HLA1 is positive and HLA2 is negative.

Embodiment 2

[0200] Muse cells (SSEA-3 positive) and non-Muse cells (SSEA-3 negative) were studied by immunocytochemical staining using an anti-human HLA-ABC antibody (eBioscience) (secondary antibody using an anti-mouse IgG antibody (labeled using Alexa568) ) in the expression of HLA class I. The results are as follows image 3 shown. Such as image 3 As shown, the expression of HLA class I can be seen in both Muse cells and non-Muse cells.

[0201] Muse cells (SSEA-3 positive) and non-Muse cells (SSEA-3 negative) were studied by immunocytochemical staining using anti-human HLA-DR antibody (eBioscience) (secondary antibody using anti-mouse IgG antibody (labeled using Alexa568) ) in the expression of HLA class II. The results are as Figure 4 shown. Such as Figure 4 As shown, there was no expression of HLA class II in both Muse cells and non-Muse cells.

[0202] Figure 5 is a graph showing the non-specific reaction in the above test. Such as Figure 5 As shown, no fluorescence w...

Embodiment 3

[0203] Example 3 Studying the Immunosuppressive Effect of Muse Cells by Lymphocyte Stimulation Test

[0204] Whether Muse cells have an immunosuppressive effect can be confirmed by an induction test of dendritic cells. That is, monocytes were isolated from human peripheral blood, co-cultured with Muse cells, and whether dendritic cells were induced by monocytes was examined.

[0205] Using Muse cells isolated from human fibroblasts with SSEA-3 positive as an indicator (hereinafter referred to as Muse cells), SSEA-3 negative cells in human fibroblasts (hereinafter referred to as non-Muse cells), and peripheral blood human monocytes.

[0206] For the medium, α-MEM (+10% FBS, 2mM L-glutamine, kanamycin) was used for Muse and non-Muse cells isolated from human fibroblasts, RPMI-1640 (+10% FBS) for human monocytes, RPMI-1640 (+10% FBS, 2mM L-glutamine, 2mM sodium pyruvate, 40ng / mL GM-CSF, 20ng / mL IL-4, kanamycin) For dendritic cell induction.

[0207] The detailed method is as ...

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Abstract

The purpose of the present invention is to provide a composition for allotransplantation cell therapy, said composition containing a pluripotent stem cell. A composition for allotransplantation cell therapy, said composition containing a SSEA-3 positive pluripotent stem cell which is capable of being isolated from body tissue and which does not express HLA class II antigens.

Description

technical field [0001] The present invention relates to a composition for cell therapy for allotransplantation containing SSEA-3 positive pluripotent stem cells derived from living tissue. Background technique [0002] In recent years, adult stem cells or tissue stem cells that can contribute to tissue regeneration have attracted attention. [0003] As cells with differentiation ability obtained from adults, for example, bone marrow mesenchymal cell fractions obtained by applying differentiation induction and having the ability to differentiate into various cells such as bone, cartilage, fat cells, nerve cells, and skeletal muscle have been reported. (MSC: Bone marrow stromal cell) (see Non-Patent Documents 1 and 2). However, the bone marrow mesenchymal cell fraction is a cell population containing multiple cell types, and the differentiation efficiency when induced is not high. It can be speculated that some cells in MSCs are responsible for differentiation, but the above...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P37/06C12N5/0735C12N5/0775A61K35/545
CPCA61K35/545A61P37/06C12N5/0607
Inventor 出泽真理吉田正顺黑田康胜
Owner 克里奥公司
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