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-8961955-B2
Publication Date
2015-02-24
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
The invention provides compositions, materials, and methods for maintaining and propagating mammalian mesenchymal stem cells (MSCs) in an undifferentiated state without the use of feeder cells. This is achieved by culturing MSCs on a three-dimensional extracellular matrix (ECM) derived from marrow stromal cells, which mimics the MSC niche in bone marrow. The ECM promotes self-renewal and retention of multipotentiality, preventing "spontaneous" differentiation towards the osteoblast lineage while preserving the ability to differentiate in response to appropriate signals.
The problem addressed by the invention is that MSCs are rare in bone marrow and difficult to expand ex vivo using traditional cell culture methods. Under standard conditions, MSCs tend to lose their stem cell properties and spontaneously differentiate, limiting their practical and therapeutic use. This loss of stemness suggests that current culture systems lack critical components of the marrow microenvironment necessary for maintaining MSC self-renewal and multipotentiality.
The invention demonstrates that culturing MSCs on a cell-free, marrow stromal cell-derived ECM provides a specialized microenvironment or niche that retains MSC properties by promoting symmetric division to produce identical daughter cells and restraining differentiation. This ECM sequesters pro-differentiating factors such as BMP2, limiting spontaneous osteoblastogenesis, and sustains high levels of functional MSCs capable of forming bone and hematopoietic marrow tissue upon transplantation. Thus, the invention offers a system for expanding functional MSCs ex vivo for various applications.
Claims Coverage
The patent contains two independent claims focusing on a bone forming composition and a method of generating bone, incorporating key inventive features related to the extracellular matrix and transplantation vehicle.
Bone forming composition comprising MSCs cultured on marrow stromal cell derived ECM and a ceramic transplantation vehicle
A composition comprising undifferentiated mammalian mesenchymal stem cells cultured on and then removed from a three-dimensional marrow stromal cell derived extracellular matrix containing type I, II, and V collagens, syndecan-1, fibronectin, decorin, biglycan, perlecan, and laminin, combined with a transplantation vehicle comprising a ceramic powder.
Method of generating bone using MSCs cultured on marrow stromal cell derived ECM with transplantation vehicle administration
A method involving obtaining mammalian mesenchymal stem cells, producing a bone forming composition by culturing these cells in vitro on a three-dimensional marrow stromal cell derived extracellular matrix comprising specified ECM components, isolating undifferentiated MSCs, combining them with a ceramic powder transplantation vehicle, and administering this composition to a patient.
The independent claims principally cover the composition of undifferentiated MSCs expanded on a specialized marrow stromal cell derived ECM combined with a ceramic transplantation vehicle, and a method of generating bone in a patient using these cultured MSCs with the extracellular matrix and transplantation vehicle. The inventive features emphasize the use of a three-dimensional marrow stromal cell derived ECM with specified components and a ceramic powder vehicle for clinical application.
Stated Advantages
The ECM promotes self-renewal and maintains MSCs in an undifferentiated state, preventing spontaneous differentiation.
Culturing MSCs on the marrow stromal cell derived ECM increases the yield of functional MSCs with enhanced bone and hematopoietic tissue formation upon transplantation.
The ECM sequesters BMP2, limiting premature osteoblast differentiation and preserving multipotentiality.
The three-dimensional ECM supports symmetric MSC division and retention of stem cell properties, unlike standard plastic culture surfaces or simple collagen/fibronectin coatings.
Documented Applications
Expansion of functional mammalian mesenchymal stem cells ex vivo for therapeutic use.
Use of MSCs cultured on the marrow stromal cell derived ECM for generating bone and hematopoietic marrow tissue in vivo after transplantation.
Treatment of bone conditions requiring bone formation such as fracture, delayed unions, non-unions, distraction osteogenesis, osteotomy, osseointegration, and osteoarthritis in mammals including humans.
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