Methods for producing fibroadipogenic progenitor cells
Inventors
Kyba, Michael S. • Goloviznina, Natalya • Xie, Ning • Dandapat, Abhijit
Assignees
University of Minnesota System
Publication Number
US-12221629-B2
Publication Date
2025-02-11
Expiration Date
2041-06-14
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
The present disclosure provides methods of producing a preparation of fibroadipogenic progenitors (FAPs) from a cell mixture. In certain embodiments, the present disclosure provides a method of producing a preparation of human FAPs from a skeletal muscle biopsy sample for later use.
Core Innovation
The invention provides methods for producing a preparation of fibroadipogenic progenitors (FAPs) from a cell mixture, specifically using CD73 as a single positive selection marker. The method involves contacting a cell mixture, dissociated from human skeletal muscle tissue, with an anti-CD73 agent having affinity for CD73, and then separating the anti-CD73 agent labeled cells to obtain a preparation of FAPs. The process avoids culturing the cell mixture under culturing conditions before selection and does not utilize additional positive co-staining selection agents.
This invention addresses the challenge of isolating and identifying FAPs in human skeletal muscle, which has been difficult due to the lack of robust and efficient marker-based methods. While FAP populations have been well-characterized and isolated in mice using combinations of cell surface markers, reliable isolation techniques for human FAPs were lacking. The method disclosed enables efficient isolation of high-quality FAPs suitable for basic research, translational applications, and potential clinical use.
The patent further provides methods to utilize the isolated FAPs for producing differentiated cell types such as adipocytes and fibroblasts, by culturing the FAPs under specific adipogenic or fibrogenic conditions. It also describes applications involving the use of FAPs or their differentiated progeny in treating muscle disorders and for compound testing in research environments.
Claims Coverage
The patent claims coverage for a method with a set of inventive features focusing on the production and preparation of fibroadipogenic progenitors (FAPs) from human skeletal muscle tissue.
Production of FAPs using anti-CD73 agent without additional positive selection
A method in which a cell mixture, freshly dissociated from human skeletal muscle tissue and not cultured prior to selection, is contacted with an anti-CD73 agent having affinity for CD73 (optionally tagged) and the anti-CD73 labeled cells are separated to produce a preparation of FAPs. No additional positive co-staining selection agent is used, and myogenic cells are excluded by this process.
The main inventive feature claimed is the selective production and isolation of FAPs from unprocessed, non-cultured human skeletal muscle tissue using only an anti-CD73 agent for positive selection, excluding myogenic cells without additional positive co-staining agents.
Stated Advantages
Provides a simple and robust method for identifying and isolating FAPs from fresh human skeletal muscle.
Enables the production of high purity preparations of FAPs suitable for downstream applications, including basic research and clinical use.
Avoids the need for culturing cells prior to selection, reducing the risk of marker expression changes and contamination.
Documented Applications
Producing a preparation of human FAPs from a skeletal muscle biopsy sample for later use.
Generating adipocytes by culturing isolated FAPs under adipogenic conditions.
Generating fibroblasts by culturing isolated FAPs under fibrogenic conditions.
Treating muscle disorders, including muscular dystrophy, by administering preparations of FAPs or differentiated cells to a subject in need.
Testing compounds for efficacy in inhibiting fibrosis, inhibiting fat cell differentiation, or promoting muscle homeostasis, repair or myogenesis using cultured FAP preparations.
Interested in licensing this patent?