Chemically defined serum replacements for cell culture
Inventors
Assignees
Montana State University Bozeman
Publication Number
US-11692167-B2
Publication Date
2023-07-04
Expiration Date
2042-08-02
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Abstract
Disclosures herein are directed to chemically defined animal-derived component free supplements designed for individual cell types that supports the ex vivo growth of cells as well or better than serum, in chemically defined conditions.
Core Innovation
The invention relates to chemically defined animal-derived component free supplements specifically formulated for individual cell types to fully replace serum in ex vivo cell culture media. These supplements are designed to support the growth of cells as well or better than serum, and are formulated to be both chemically defined and free of any animal-derived products, including albumin.
Traditional use of sera and serum albumins in cell culture introduces significant variability and risk due to unknown and variable components, and potential for contamination with toxins, drugs, prions, and viral particles. Such contamination can negatively affect cell viability and phenotype, making validation processes expensive and sometimes impossible. This invention addresses the recognized problems by providing defined formulations composed only of precisely known components, each tailored to the extracellular needs of specific cell types.
The core innovation includes supplements containing combinations of specific lipids, trace elements, hydrophobic vitamins, antioxidants, and growth factors, formulated without animal-derived components. The resulting supplements are customized for various cell types, such as induced pluripotent stem cells (iPSCs), human foreskin fibroblasts (HFF), human embryonic kidney (HEK) cells, and smooth muscle cells (SMCs), to ensure optimal growth, proliferation, and health in cell culture under chemically defined, contaminant-free conditions.
Claims Coverage
There are three main inventive features derived from the independent claims of the patent.
Chemically defined, animal-derived product free media supplement with specific lipid and vitamin composition
A chemically defined media supplement that comprises: - Ascorbyl palmitate - Docosahexanoic acid - Palmitoleic acid - Cholesterol sulfate - At least two components selected from the group consisting of ascorbic acid, silver, vitamin A, vitamin D3, and vitamin K The supplement is explicitly animal-derived product free.
Chemically defined media supplement with ITSE, trace elements, growth factors, and omega-3 fatty acids
A media supplement comprising: - ITSE (Insulin, transferrin, selenium, ethanolamine) - Trace Elements A - Alpha linolenic acid - Fibroblast growth factor - Insulin-like growth factor - Oleic acid - Docosahexanoic acid - Vitamin D3 The supplement is animal-derived product free.
Chemically defined media supplement with ITSE, trace elements, lipids, and growth factors for tailored cell support
A media supplement comprising: - ITSE (Insulin, transferrin, selenium, ethanolamine) - Trace Elements A - Alpha linolenic acid - Fibroblast growth factor - Insulin-like growth factor - Cholesterol sulfate - Palmitoleic acid The supplement is animal-derived product free.
The inventive features collectively cover compositions of chemically defined, animal-derived product free supplements with specifically stated combinations of lipids, vitamins, trace elements, and growth factors, optimized for cell type-specific support in cell culture media.
Stated Advantages
Provides cell culture supplements that are both chemically defined and free from animal-derived products, thereby eliminating the risk of contamination from animal sources.
Reduces variability and inconsistency seen with serum or serum albumin by using only precisely identified and quantified components.
Improves the safety of cell cultures and resulting cell products for therapeutic use or human consumption by preventing potential contamination with toxins, drugs, prions, or viruses.
Allows for formulation of cell type-specific supplements that can support ex vivo cell growth as well or better than serum under chemically defined conditions.
Documented Applications
Supplementation of culture media for induced pluripotent stem cells (iPSCs), human foreskin fibroblasts (HFF), human embryonic kidney (HEK) cells, and smooth muscle cells (SMCs).
Production of cells for therapeutic applications, including stem cell transplantation and production of organs (e.g., larynx, skin) for transplant.
Production of other cell types for vaccine production, protein production, and medical research and testing.
Cultivation of animal cells (e.g., human cells, chicken cells, monkey cells, bovine cells, etc.) under serum-free, chemically defined conditions.
Cultivation of cells that are capable of producing vaccines, viruses, viral particles, viral proteins, nucleic acids, or viral fragments.
Preferential expansion of subpopulations of cells (such as mononuclear cell subpopulations) by adjusting nutrient molar ratios.
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