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Publication Number
US-10653724-B2
Publication Date
2020-05-19
Expiration Date
Abstract
Isolated extracellular matrix from marrow adherent stromal cells and their descendents, which stimulates the growth and survival of a number of different neural cell types, is provided, along with methods for preparation and uses.
Core Innovation
The invention relates to an isolated extracellular matrix (ECM) produced by culturing cells on a culture surface for a predetermined period of time and removing the cells from the culture surface. The source cells are marrow adherent stromal cells (MASCs)/mesenchymal stem cells and differentiation-restricted descendents (DRCs), and the DRCs are generated by Notch intracellular domain (NICD) transfection and selection.
The isolated ECM supports neural regeneration by supporting neuron survival and growth, including neurite outgrowth, as well as supporting glial growth, including astrocytes and oligodendrocytes. The document states that the ECM can be used in vitro and as a transplantable substrate for treating neural degeneration/disorders in both the central nervous system and peripheral nervous system.
A hypothesized mechanism is provided in which the ECM functions as a growth substrate and provides matrix-bound factors that support neural cells. The document also describes comparison results between MASC-ECM and DRC-ECM, including comparisons versus poly-D-lysine (PDL), and states that observed effects are not due to enhanced initial adhesion.
Claims Coverage
The claim set includes three independent claims covering an isolated ECM, a method for stimulating neural cell growth, and a method for treating neural degeneration or disorder. The inventive features center on three claim types.
Notch intracellular domain-derived isolated ECM supporting neuron growth
An isolated extracellular matrix (ECM) produced by culturing cells on a culture surface for a predetermined period of time and removing the cells from the culture surface, wherein the cells are obtained by transfecting marrow adherent stromal cells with a nucleic acid comprising sequences encoding a Notch intracellular domain, culturing in the presence of a selective agent to select cells comprising the nucleic acid, and further culturing in the absence of the selective agent, wherein the isolated ECM supports growth of neurons to a greater extent than an ECM produced by a marrow adherent stromal cell.
Stimulating neural cell growth on Notch intracellular domain-derived isolated ECM
A method for stimulating the growth of neural cells by culturing a population of neural cells on an isolated extracellular matrix (ECM) produced by culturing cells on a culture surface for a predetermined period of time and removing the cells from the culture surface, wherein the cells are obtained by transfecting marrow adherent stromal cells with a nucleic acid comprising sequences encoding a Notch intracellular domain, culturing in the presence of a selective agent to select cells comprising the nucleic acid, and further culturing in the absence of the selective agent.
Treating neural degeneration or disorder with transplantable Notch intracellular domain-derived isolated ECM
A method for treating neural degeneration or disorder in a subject by transplanting to a site in the subject an isolated extracellular matrix (ECM) produced by culturing cells on a culture surface for a predetermined period of time, removing the cells from the culture surface, and collecting the ECM from the culture surface, wherein the cells are obtained by transfecting marrow adherent stromal cells with a nucleic acid comprising sequences encoding a Notch intracellular domain, culturing in the presence of a selective agent to select cells comprising the nucleic acid, and further culturing in the absence of the selective agent.
The independent claims cover an isolated ECM produced from marrow adherent stromal cells via Notch intracellular domain transfection and selective agent selection, a method for stimulating neural cell growth on that ECM, and a method for treating neural degeneration or disorder by transplanting the ECM.
Stated Advantages
The isolated ECM supports growth of neurons to a greater extent than an ECM produced by a marrow adherent stromal cell.
Documented Applications
Supporting neural regeneration in vitro, including neuron survival and growth and neurite outgrowth, and glial growth including astrocytes and oligodendrocytes.
Transplantable use as a substrate for treating neural degeneration/disorders in a subject.
Central nervous system transplant application.
Peripheral nervous system transplant application.
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