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Ovarian surface epithelium cell 3D-EMT immunocompetence preparation and preparation and application

A 3D-EMT, immune activity technology, applied in the field of pluripotent immune activity preparations and preparation and application, can solve the problems of immune diseases, differentiation maladaptation, immune disorders, etc.

Active Publication Date: 2019-12-31
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although exogenous thymus-related tissue or cell transplantation is expected to correct this immune deficiency to a certain extent, due to the inadaptability of exogenous thymic epithelial cells (TECs) to the development and differentiation of lymphoid progenitor cells in the host's intrinsic tissue, Selection of exported lymphocytes may lead to body-specific autoimmune diseases or other immune disorders, thereby inducing human leukocyte antigen (Human leukocyte antigen, HLA) / major histocompatibility complex (Majorhistocompatibility complex, MHC) incompatibility events happened

Method used

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  • Ovarian surface epithelium cell 3D-EMT immunocompetence preparation and preparation and application
  • Ovarian surface epithelium cell 3D-EMT immunocompetence preparation and preparation and application
  • Ovarian surface epithelium cell 3D-EMT immunocompetence preparation and preparation and application

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Embodiment 1

[0027] Embodiment 1. A 3D-EMT immune active preparation of ovarian surface epithelial cells is a 3D-EMT microsphere active preparation of ovarian surface epithelial cells modified by 160Gy X-ray irradiation in a 2ml cryopreservation tube, 3D -The number of EMT microspheres is 5×10 3 Each microsphere, with a diameter of 195±25 μm, was frozen in human AB plasma with a total volume of 1 ml and stored in liquid nitrogen at -196°C until use.

Embodiment 2

[0028] Embodiment 2. The preparation of the 3D-EMT immune active preparation of ovarian surface epithelial cells: its operation is to be divided into following five steps preparations under sterile conditions: (1) separate ovarian surface epithelial cells; (2) ovarian surface epithelial cells (3) Ovarian surface epithelial cells were induced to transform into 3D-EMT microspheres in a serum-free dynamic suspension culture system, and the diameter of the microspheres was 195±25μm; (4) 3D-EMT microspheres received 160Gy X-rays Irradiation modification; (5) preparation of radiation-modified 3D-EMT microsphere immunoactive preparations: collect 3D-EMT microspheres and put them into 2ml cryopreservation vials, the number of each 3D-EMT microspheres is 5×10 3 Each, the diameter of microspheres is 195±25μm, the cryopreservation medium is human AB plasma, the total volume is 1ml, stored in liquid nitrogen at -196°C, and used directly after thawing.

[0029] The preparation procedure is...

Embodiment 3

[0035] Example 3. Identification of the pluripotent type of the 3D-EMT immune active preparation of ovarian surface epithelial cells: the ovarian surface epithelial cells were adhered to the wall for conventional culture (Control), and the adherent conventional cultured cells were modified by irradiation (2D-CB) and 3D-EMT spheres modified by irradiation (3D-EMT) these three kinds of cells through the sequencing of the transcriptome and bioinformatics analysis, the clustering heat map of the main differentially expressed genes of the three groups of Control, 2D-CB and 3D-ETSB comparison (see attached image 3 ).

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Abstract

The invention relates to an ovarian surface epithelium cell 3D-EMT immunocompetence preparation and preparation and an application. An ovarian surface epithelium cell 3D-EMT microsphere activity preparation subjected to 160Gy X ray irradiation modification is preserved in a frozen-storage pipe of 2ml, the number of microspheres is 5*10<3>, the diameter of the microspheres is 195+ / -25 [mu]m, frozen-storage liquid is human AB plasma, the total volume is 1ml, and the frozen-storage liquid is preserved in liquid nitrogen of minus 196 DEG C. A preparation method comprises five steps under the bacteria-free condition: 1, separating ovarian surface epithelium cells; 2, performing adherence enlarged culture on the ovarian surface epithelium cells; 3, inducing the ovarian surface epithelium cells in a serum-free dynamic suspension culture system to be converted into 3D-EMT microspheres, wherein the diameter of the microspheres is 195+ / -25 [mu]m; 4, enabling the 3D-EMT microspheres to receive Xray irradiation modification; and 5, preparing the 3D-EMT microsphere immunocompetence preparation. The preparation disclosed by the invention creates bran-new medicinal preparation application for related diseases of AIDS, malignant tumors at progressive stage, chronic infection, elderly immune hypofunction and the like caused by immunosenescence / immunodeficiency.

Description

technical field [0001] The present invention relates to biomedical engineering technology, especially a method for inducing ovarian surface epithelial cells to undergo epithelial-mesenchymal transition (Epithelial-mesenchymal transition, EMT) and mesenchymal-epithelial transition (Mesenchymal-epithelial transition, MET) by using a serum-free dynamic suspension method. The prepared 3D-EMT microspheres were modified by radiation irradiation to prepare multipotent immune active preparations and their preparation and application. Background technique [0002] Ovarian surface epithelium cell (OSEC) is a type of cell with indeterminate phenotype and indistinct differentiation characteristics, which can be stimulated by environmental changes to differentiate in multiple directions. It is a single layer of cuboids coated on the surface of the ovary Or rectangular, epithelial cells with mesenchymal properties. Previously known as the "germinal epithelium," the surface epithelium of ...

Claims

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Application Information

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IPC IPC(8): A61K35/54A61K9/50A61P31/18A61P35/00A61P31/00A61P37/04C12N5/0775C12N5/071C12N13/00
CPCA61K35/54A61K9/50A61P31/18A61P35/00A61P31/00A61P37/04C12N5/0668C12N5/0625C12N13/00C12N2509/00
Inventor 张艳娜陈县城
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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