Cell culture media and methods for generating human alveolar macrophage-like cells
Inventors
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Assignees
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Texas Biomedical Research InstituteTexas Biomedical Research Institute (Texas Biomed) is a leading nonprofit biomedical research organization with more than 80 years of experience supporting the development of medical countermeasures for emerging and high-consequence infectious diseases. Texas Biomed provides comprehensive in-vitro and in-vivo preclinical research services supporting the discovery, development, and evaluation of vaccines, therapeutics, diagnostics, and other medical countermeasures from early discovery through preclinical proof-of-concept, efficacy, and translational research. Studies can be conducted under GLP or GLP-like quality frameworks, as appropriate to program stage and sponsor requirements, with integrated quality assurance and study compliance support.
The Institute operates state-of-the-art ABSL-2, ABSL-3, as well as the only privately operated BSL-4 research facilities, together with the Southwest National Primate Research Center, enabling studies in small animal and nonhuman primate models under high-containment conditions.
Texas Biomed has extensive expertise in virus propagation and characterization, antiviral and neutralization assays, molecular diagnostics, immunology, pathology, bioinformatics, animal model development, natural history studies, efficacy testing, pharmacology, and biomarker evaluation across a broad range of viral pathogens, including filoviruses, henipaviruses, arenaviruses, alphaviruses, flaviviruses, coronaviruses, hantaviruses, and other emerging and biodefense-relevant viruses. Pathogen expertise includes, but is not limited to, Ebola virus (EBOV), Sudan virus (SUDV), Bundibugyo virus (BDBV), Marburg virus (MARV), Nipah virus (NiV), Lassa virus (LASV), Junín virus (JUNV), Machupo virus (MACV), Western equine encephalitis virus (WEEV), Japanese encephalitis virus (JEV), West Nile virus (WNV), Dengue virus (DENV) and SARS-CoV-2.
With decades of experience supporting Animal Rule programs and infectious disease research, Texas Biomed delivers integrated, high-quality preclinical solutions for government, industry, and academic partners developing medical countermeasures for BSL-2, BSL-3, and BSL-4 pathogens.
Texas Biomedical Research Institute (Texas Biomed) is a leading nonprofit biomedical research organization with more than 80 years of experience supporting the development of medical countermeasures for emerging and high-consequence infectious diseases. Texas Biomed provides comprehensive in-vitro and in-vivo preclinical research services supporting the discovery, development, and evaluation of vaccines, therapeutics, diagnostics, and other medical countermeasures from early discovery through preclinical proof-of-concept, efficacy, and translational research. Studies can be conducted under GLP or GLP-like quality frameworks, as appropriate to program stage and sponsor requirements, with integrated quality assurance and study compliance support. The Institute operates state-of-the-art ABSL-2, ABSL-3, as well as the only privately operated BSL-4 research facilities, together with the Southwest National Primate Research Center, enabling studies in small animal and nonhuman primate models under high-containment conditions. Texas Biomed has extensive expertise in virus propagation and characterization, antiviral and neutralization assays, molecular diagnostics, immunology, pathology, bioinformatics, animal model development, natural history studies, efficacy testing, pharmacology, and biomarker evaluation across a broad range of viral pathogens, including filoviruses, henipaviruses, arenaviruses, alphaviruses, flaviviruses, coronaviruses, hantaviruses, and other emerging and biodefense-relevant viruses. Pathogen expertise includes, but is not limited to, Ebola virus (EBOV), Sudan virus (SUDV), Bundibugyo virus (BDBV), Marburg virus (MARV), Nipah virus (NiV), Lassa virus (LASV), Junín virus (JUNV), Machupo virus (MACV), Western equine encephalitis virus (WEEV), Japanese encephalitis virus (JEV), West Nile virus (WNV), Dengue virus (DENV) and SARS-CoV-2. With decades of experience supporting Animal Rule programs and infectious disease research, Texas Biomed delivers integrated, high-quality preclinical solutions for government, industry, and academic partners developing medical countermeasures for BSL-2, BSL-3, and BSL-4 pathogens.
Publication Number
US-12173326-B2
Publication Date
2024-12-24
Expiration Date
2042-03-30
Abstract
Provided here methods of generating human alveolar macrophage-like cells in-vitro from blood-derived monocytes by culturing them in a cell culture medium containing a mixture of a surfactant and two or more cytokines. This mixture can contain calfactant, granulocyte-macrophage colony-stimulating factor, transforming growth factor beta, and interleukin-10.
Core Innovation
The invention provides methods for generating human alveolar macrophage-like (AML) cells in vitro from blood-derived monocytes by culturing them in a specialized cell culture medium containing a mixture of a surfactant and two or more cytokines. This mixture can specifically include calfactant, granulocyte-macrophage colony-stimulating factor (GM-CSF), transforming growth factor beta (TGF-β), and interleukin-10 (IL-10), either individually or in a defined combination. The invention further includes a cell culture medium tailored with these components to reliably convert blood-derived monocytes into cells exhibiting human alveolar macrophage phenotype and function.
The problem addressed by this invention is the significant difficulty in obtaining authentic human alveolar macrophages (HAM) for research due to limited access from donors, the expense and labor involved in bronchoalveolar lavage, and the rapid phenotypic changes that occur during in vitro culture. These challenges restrict the ability to study HAM roles in lung diseases and limit the availability of these cells for use in commercial cell assays, diagnostics, drug screening, and basic research.
By providing a cell culture system and defined medium capable of converting easily accessible blood-derived monocytes into HAM-like cells, the invention offers an alternative to directly harvesting HAM from lung tissues or bronchoalveolar lavage. The system mimics the alveolar environment, enabling the expansion and use of HAM-like cells for various applications while overcoming the limitations of traditional isolation methods.
Claims Coverage
There are two independent claims in this patent, each describing distinct inventive features relating to the generation of human alveolar macrophage-like cells from blood-derived monocytes using specific cell culture media and conditions.
Method for generating human alveolar macrophage-like cells using surfactant and cytokine mixture
The method comprises culturing a population of human blood-derived monocytes in a cell culture medium containing a mixture of a surfactant and two or more cytokines. The resulting human alveolar macrophage-like (AML) cells have increased expression of one or more of the following markers as compared to the starting monocytes: MRC1 (CD206), MARCO, MERTK, CES1, CD170, MCL1, CCL18, CXCL3, DUSP1, CXCL5, PU.1, H3K4mel, CD200R, CD11c, CD163, MCEMP1, and/or decreased expression of one or more of CD11b, CD36, MMP7, MMP9, H3K4me3.
Method for generating human alveolar macrophage-like cells using a precise combination of calfactant, GM-CSF, TGF-β, and IL-10
This method involves culturing human blood-derived monocytes in a cell culture medium that specifically includes a mixture of calfactant, granulocyte-macrophage colony-stimulating factor, transforming growth factor beta, and interleukin 10 to generate AML cells.
The inventive features cover methods of generating AML cells from blood-derived monocytes using a medium containing a mixture of surfactant and cytokines, as well as a specific medium comprising calfactant, GM-CSF, TGF-β, and IL-10. Both methods are directed to obtaining AML cells with defined marker expression profiles from readily available blood-derived monocytes.
Stated Advantages
Provides a cost-effective and accessible method for generating, culturing, and maintaining human alveolar macrophage-like cells from blood-derived monocytes, enabling broader and easier study of HAM biology.
Enables generation of AML cells that exhibit phenotype and function similar to authentic human alveolar macrophages, which are otherwise difficult to obtain.
Overcomes phenotypic instability and scarcity issues associated with direct isolation of HAM from human donors.
Facilitates large-scale production of HAM-like cells for use in commercial cell assays and research applications.
Documented Applications
Studying pharmacology, toxicology, drug development, drug delivery, drug metabolism, drug-drug interaction, drug bioavailability, and drug clearance.
Examining multi-organ interactions, diagnostics, therapeutics, and nutritional applications.
Researching physiology of the lung, respiratory disease models, mechanisms of disease, and etiology of disease in the respiratory system.
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