Maintenance and propagation of mesenchymal stem cells
Inventors
Chen, Xiao-Dong • Jilka, Robert L.
Assignees
US Department of Veterans Affairs • University of Arkansas at Little Rock
Publication Number
US-9873862-B2
Publication Date
2018-01-23
Expiration Date
2027-01-22
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Abstract
Various embodiments of the present invention include compositions, materials and methods for maintaining and propagating mammalian mesenchymal stem cells in an undifferentiated state in the absence of feeder cells and applications of the same.
Core Innovation
Various embodiments of the present invention include compositions, materials, and methods for maintaining and propagating mammalian mesenchymal stem cells (MSCs) in an undifferentiated state in the absence of feeder cells, utilizing an extracellular matrix (ECM) derived from marrow stromal cells. This ECM specifically promotes self-renewal of MSCs and retention of their multipotentiality, facilitating their expansion and maintenance ex vivo.
The problem being solved arises from the extreme rarity of MSCs in the bone marrow and the difficulty of expanding them ex vivo while retaining their stem cell properties. Traditional culture conditions result in loss of these properties, impairing practical and therapeutic use of MSCs. The invention addresses the need for a specialized microenvironment or niche that supports MSC self-renewal and multipotentiality, which is lacking in standard in vitro culture systems.
The invention demonstrates that culturing MSCs on a three-dimensional ECM produced by marrow stromal cells reconstitutes this niche and supports MSC self-renewal by promoting symmetric cell division, while restraining spontaneous differentiation. This 3D stromal cell-derived ECM preserves MSC multipotentiality and functional capabilities, such as forming complete bone-like structures in vivo, which are not maintained on tissue culture plastic or simpler 2D matrices. Moreover, the ECM sequesters pro-differentiating factors like BMP2, contributing to maintenance of the undifferentiated state.
Claims Coverage
The claims include one independent method claim that covers maintaining mammalian mesenchymal stem cells in culture in an undifferentiated state using a specific extracellular matrix comprising defined components.
Method of maintaining MSCs in an undifferentiated state using a specified extracellular matrix
A method of maintaining mammalian mesenchymal stem cells in culture in an undifferentiated state by culturing the cells in the presence of an extracellular matrix that comprises type I collagen, type III collagen, type V collagen, syndecan-1, fibronectin, decorin, biglycan, perlecan, and laminin.
The claims focus on the use of a marrow stromal cell-derived, three-dimensional extracellular matrix containing specific collagen types and proteoglycans to preserve mammalian mesenchymal stem cells in an undifferentiated and multipotent state during culture, enabling their maintenance and propagation without feeder cells.
Stated Advantages
The marrow stromal cell-derived ECM promotes self-renewal of MSCs and retention of their multipotentiality.
The ECM restrains spontaneous differentiation of MSCs toward the osteoblast lineage under culture conditions.
MSCs expanded on the stromal cell-derived ECM exhibit enhanced capacity to form bone and hematopoietic marrow in vivo compared to those expanded on plastic or simpler matrices.
The 3D ECM sequesters BMP2 and possibly other pro-differentiating factors, contributing to maintenance of MSC stemness.
The system permits expansion of functional MSCs ex vivo for practical applications and enables study of the effects of aging or hormonal changes on MSC function.
Documented Applications
Expansion of functional mammalian mesenchymal stem cells ex vivo for therapeutic and practical uses.
Generation of bone-forming compositions comprising MSCs cultured on stromal cell-derived ECM combined with transplantation vehicles such as hydroxyapatite/tricalcium phosphate ceramic powders.
Treatment of conditions requiring bone formation, remodeling, or repair including fractures, delayed unions, non-unions, distraction osteogenesis, osteotomy, osseointegration, and osteoarthritis.
Use in identification of specific ECM component effects on MSC behavior and studying pathological conditions such as osteoporosis related to aging or hormonal changes.
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