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Method for selecting high-efficacy stem cell by using downregulation in expression or activity of socs

A technology of activity inhibitors and mesenchymal stem cells, which is applied in the field of selecting high-efficiency stem cells by inhibiting the expression or activity of SOCS, can solve the problems of unremarkable results, lack of culture methods or programs, etc., and achieve the effect of high therapeutic possibility

Pending Publication Date: 2019-07-30
细胞生命有限公司
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AI Technical Summary

Problems solved by technology

The immunosuppressive properties of MSCs are known, and although most phase I clinical trials have not shown biostability issues, less outstanding results have been obtained in later clinical stages
[0008] Furthermore, due to the lack of culture methods or protocols for MSC standardization, phenotypic and functional diversity was observed even when MSCs were isolated from the same tissue from the same recipient

Method used

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  • Method for selecting high-efficacy stem cell by using downregulation in expression or activity of socs
  • Method for selecting high-efficacy stem cell by using downregulation in expression or activity of socs
  • Method for selecting high-efficacy stem cell by using downregulation in expression or activity of socs

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1: experimental method

[0071] 1-1. Isolation and culture of human tissue-derived MSCs

[0072] This experiment was approved by the Institutional Review Board (IRB) of Samsung Medical Center (IRB No. 2011-10-134), and all samples were obtained with informed consent. MSCs are isolated by conventional known methods. Separated cells in 2 x 10 3 cells / cm 2 The density was inoculated in Dulbecco's modified Eagle's medium (DMEM; Invitrogen-Gibco, Rockville, MD) containing 10% fetal bovine serum (FBS, Invitrogen-Gibco) and 100U / mL penicillin / streptomycin (Invitrogen -Gibco), and at 37°C and 5% CO 2 cultivated under conditions.

[0073] 1-2. shRNA transfection

[0074]To inhibit SOCS1 expression, adipose tissue-derived MSCs (AT-MSCs) were treated with adenovirus expressing SOCS1 shRNA and red fluorescent protein (RFP). Specifically, the shRNA sequence targeting SOCS1 was cloned into the shuttle vector (pO6A5-U6-mPGK-TagRFP) containing the human U6 promoter ...

Embodiment 2

[0079] Embodiment 2: experimental result

[0080] 2-1. Inhibition of SOCS expression

[0081] Suppression of SOCS expression in MSCs was confirmed by transduction of lipid tissue-derived MSCs (AT-MSCs) with adenovirus expressing shRNA targeting SOCS1. The result is as Figure 1a and 1b As shown, from the RFP tag ( Figure 1a ) and Western blot ( Figure 1b ) results can confirm that the expression of SOCS1 protein in MSCs is significantly inhibited.

[0082] 2-2. Evaluation of in vivo immunosuppressive activity of SOCS-suppressed MSCs

[0083] It was confirmed whether the SOCS-suppressed MSCs obtained in Example 2-1 actually exhibit immunosuppressive properties in the GVHD animal model.

[0084] The result is as figure 2 As shown, it can be seen that the shRNA-inhibited MSC (red line) treated group with SOCS has significantly improved survival (immunosuppressive properties) compared to the control.

[0085] 2-3. Discovery of other immunosuppressive biomarkers in SOCS-...

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Abstract

The present invention relates to a method for selecting high-efficacy mesenchymal stem cells for use in treating an immune disease, the method comprising a step of measuring a level of a particular immunosuppressive biomarker in mesenchymal stem cells in which the expression or activity of suppressor of cytokine signaling (SOCS) is downregulated, high-efficacy mesenchymal stem cells selected by the method, and a method for treating an immune disease by using the high-efficacy mesenchymal stem cells. Providing a method useful for acquiring functionally excellent mesenchymal stem cells having anability to control immune reactions for clinical treatment of various immune diseases including graft-versus-host diseases and autoimmune diseases, the present invention can find useful applicationsin the therapy of immune diseases.

Description

technical field [0001] The invention of the present application relates to a method for selecting high-efficiency mesenchymal stem cells for the treatment of immune diseases, said method comprising determining the level of specific immunosuppressive biomarkers in mesenchymal stem cells, wherein the cytokine signaling in said mesenchymal stem cells The expression or activity of transduction suppressor factor (SOCS) is inhibited, high-efficiency mesenchymal stem cells selected by the method, and a method for using the high-efficiency mesenchymal stem cells to treat immune diseases. Background technique [0002] Organ, tissue or cell transplantation can be used to save the life of patients suffering from various diseases. The allogeneic transplantation of human organs such as kidney, liver, heart, lung, pancreas, etc., tissue such as skin, and cells such as bone marrow cells is a common clinical practice for the treatment of intractable diseases such as end organ failure. solu...

Claims

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Application Information

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IPC IPC(8): G01N33/50A61K35/28
CPCA61K35/28G01N33/50C12N15/113A61P37/06G01N33/5073G01N2800/245G01N2800/7095C12N2310/14C12N2310/531G01N2500/10C12N5/0662C12N2501/999
Inventor 刘建喜李明雨金大成
Owner 细胞生命有限公司
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