Gene and protein expression properties of adherent stromal cells cultured in 3D
Inventors
Assignees
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Publication Number
US-10351910-B2
Publication Date
2019-07-16
Expiration Date
Abstract
Adherent stromal cells cultured under three dimensional conditions are provided, characterized, and distinguished from adherent stromal cells cultured under two dimensional conditions.
Core Innovation
The disclosure relates to adherent stromal cells (ASCs) produced by 3D culture, where the 3D culture comprises polyester 3D carriers, and to distinguishing these ASC-3D cells from ASC-2D cells. Biomarker panels are used at the gene and protein levels to differentiate ASCs produced by 3D culture conditions from ASCs produced by 2D culturing.
A determining step is performed by measuring the expression, or in culture medium contexts detecting or measuring expression, of one or more biomarkers in a sample of ASC, including VEGFA and additional biomarkers such as LIF, COL7A1, IL6, MMP10, MMP9, MMP11, FN1, COL15A1, TNC, or IFNA1. The measured levels are compared to levels measured in samples of ASC produced by 2D culturing or to culture medium produced from ASCs grown in 2D culturing.
The disclosure further connects the identified biomarker signatures associated with ASC produced by 3D culture comprising polyester 3D carriers to a treating step, where the ASC is administered to a subject thereby treating an autoimmune disease. An increase of at least 2-fold relative to the 2D control indicates the cell is an ASC produced by 3D culturing comprising polyester 3D carriers.
Claims Coverage
The disclosure provides two independent methods. The inventive features include identifying ASCs produced by 3D culture with polyester 3D carriers using biomarker measurements relative to 2D controls, and administering the identified ASC to treat an autoimmune disease.
Treating an autoimmune disease using 3D-culture ASC identification by biomarker expression versus 2D control
A method comprising determining whether an ASC was produced by 3D culture comprising polyester 3D carriers by measuring in a sample of ASC the expression of VEGFA and one or more selected biomarkers (LIF, COL7A1, IL6, MMP10, MMP9, MMP11, FN1, COL15A1, TNC, or IFNA1) using gene array or PCR, comparing the measured levels to levels of the same biomarker measured in a sample of ASC produced by 2D culturing (2D control), wherein an increase of at least 2-fold of any one or more of the listed biomarkers relative to the 2D control indicates the cell is an ASC produced by 3D culturing comprising polyester 3D carriers, and administering the ASC to the subject thereby treating an autoimmune disease.
Treating an autoimmune disease using VEGFA in culture medium as a 3D-culture indicator versus 2D medium control
A method comprising determining whether an adherent stromal cell was produced by three dimensional culture comprising polyester three dimensional carriers by measuring VEGFA in a culture medium produced from a sample of adherent stromal cells using antibody array or ELISA, comparing the measured levels to levels of VEGFA in culture medium produced from adherent stromal cells grown in two dimensional culturing (2D medium control), wherein an increase of at least two-fold of VEGFA relative to the 2D medium control indicates the cell is an adherent stromal cell produced by three dimensional culturing comprising polyester three dimensional carriers, and administering the ASC to the subject thereby treating an autoimmune disease.
Across the independent claims, the core claim coverage centers on identifying ASCs produced by 3D culture with polyester 3D carriers by biomarker measurement compared to 2D controls, followed by administering the identified ASC to treat an autoimmune disease. The biomarker determination is based either on a panel of gene-expression biomarkers measured by gene array or PCR, or on VEGFA detected or measured in culture medium by antibody array or ELISA.
Stated Advantages
Enables distinguishing ASCs produced by 3D culture comprising polyester 3D carriers from ASCs produced by 2D culturing using biomarker panels at the gene and protein levels.
Supports treating an autoimmune disease by administering ASCs identified as produced by 3D culturing comprising polyester 3D carriers based on biomarker increases relative to 2D controls.
Provides a biomarker-based indication where an increase of at least 2-fold, including VEGFA, relative to 2D control indicates 3D-produced ASC.
Documented Applications
Treating an autoimmune disease in a subject in need thereof by administering an ASC that is determined to have been produced by three dimensional culture comprising polyester three dimensional carriers.
Treating an autoimmune disease in a subject in need thereof using ASC that is determined to have been produced by 3D culture comprising polyester 3D carriers based on biomarker measurement versus 2D controls.
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