Marker for Neural Stem Cells
AssigneeXintela AB
InventorsEvy Lundgren Åkerlund, Katarzyna Chmielarska Masoumi
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Abstract
The present application concerns methods for detecting and isolating a population of neural stem cells (NSC) or neural progenitor cells (NPC) based on expression of the marker integrin alpha10beta1; as well as use of said population of NSC or NPC for therapy, diagnosis and prognosis of disease and damage of the CNS.
Claims83
Use of a marker comprising an integrin alpha 10 subunit expressed by a neural stem cell and/or a neural progenitor cell as a marker for mammalian neural stem cells and mammalian neural progenitor cells.
A method for identifying a mammalian neural stem cell and/or a mammalian neural progenitor cell, the method comprising the steps of:
- a) providing a sample comprising neural tissue,
- b) detecting expression of an integrin alpha10 subunit by a cell comprised in the sample of a),
- c) scoring the integrin alpha10 subunit expression of b), and
- d) identifying the mammalian neural stem cell and/or the neural progenitor cell according to the scoring in c).
A method for isolating a mammalian neural stem cell and/or a mammalian neural progenitor cell, the method comprising the steps of:
- a) providing a sample comprising neural tissue,
- b) detecting expression of an integrin alpha10 subunit by a cell comprised in the sample of a),
- c) scoring the integrin alpha10 subunit expression of b), and
- d) selecting the mammalian neural stem cell and/or the mammalian neural progenitor cell according to the scoring in c).
The method or use according to any one of the preceding claims, wherein the integrin alpha 10 subunit is expressed on the cell surface of the mammalian neural stem cell and/or neural progenitor cell.
The method or use according to any one of the preceding claims, wherein the integrin alpha 10 subunit is expressed intracellularly in a mammalian neural stem cell and/or neural progenitor cell.
The method or use according to any one of the preceding claims, wherein the integrin alpha10 is expressed as a heterodimer in combination with an integrin beta1 subunit.
The method or the use according to any one of the preceding claims, further comprising a step of contacting the sample with an antibody which specifically binds integrin alpha10 subunit prior to the detection of b).
The method or the use according to any one of the preceding claims, further comprising detecting expression of a secondary marker selected from the group consisting of Nestin, PSA-NCAM, GFAP, PDFGRα, SOX-2, CD133 (prominin-1), CD15, CD24, Musashi, EGFR, Doublecortin (DCX), Pax6, FABP7, LeX, Vimentin and GLAST.
The method according to any one of the preceding claims, wherein the method is performed in vitro.
The method according to any one of the preceding claims, wherein the neural tissue comprises a neural stem cell and/or a neural progenitor cell.
The method according to any one of the preceding claims, wherein the neural tissue is obtained or derived from brain tissue.
The method according to any one of the preceding claims wherein the neural tissue is adult brain tissue.
The method according to any one of the preceding claims, wherein the neural tissue is fetal brain tissue.
The method according to any one of the preceding claims, wherein the neural tissue is derived from the subventricular zone (SVZ) or from the subgranular zone (SGZ) or from the meninges.
The method according to any one of the preceding claims, wherein the neural tissue is selected from the group consisting of human, canine, equine, bovine, feline, murine, ovine or swine neural tissue.
The method according to any one of the preceding claims, wherein the detection of expression of an integrin alpha10 subunit by a cell is determined by flow cytometry.
The method according to any one of the preceding claims, wherein the detection of expression of an integrin alpha10 subunit by a cell is determined by measuring integrin alpha10 protein expression.
The method according to any one of the preceding claims, wherein the detection of expression of an integrin alpha10 subunit by a cell is determined by measuring integrin alpha10 mRNA expression.
The method according to any one of the preceding claims, wherein the detection of expression of an integrin alpha10 subunit by a cell is determined by a method selected from the group consisting of immunoassay, immunoprecipitation, flow cytometry, immunofluorescence, and western blot.
The method according to any one of the preceding claims, wherein the antibody used for detection of expression of an integrin alpha10 subunit by a cell is a monoclonal antibody, polyclonal antibody, a chimeric antibody, a single chain antibody or fragment thereof.
The method according to any one of the preceding claims, wherein the antibody is a monoclonal antibody.
The method according to any one of the preceding claims, wherein the antibody is a non-human antibody, a chimeric antibody, a bispecific antibody, a humanized antibody or a human antibody.
The method according to any one of the preceding claims, wherein the antibody is covalently bound to a detectable moiety, such as a detectable moiety selected from the group consisting of a fluorophore, an enzyme or a radioactive tracer or radioisotope.
The method according to any one of the preceding claims, wherein the antibody has an isotype selected from the group consisting of IgA, IgD, IgG and IgM.
The method according to any one of the preceding claims, wherein the antibody is:
- a) a monoclonal antibody, produced by the hybridoma cell line deposited at the Deutsche Sammlung von Microorganismen und Zellkulturen GmbH under the accession number DSM ACC2583;
- or
- b) an antibody which competes for binding to the same epitope as the epitope bound by the monoclonal antibody produced by the hybridoma deposited at the Deutsche Sammlung von Microorganismen und Zellkulturen GmbH under the accession number DSM ACC2583;
- or
- c) a fragment of a) or b), wherein said fragment is capable of binding specifically to the extracellular I-domain of the integrin alpha 10 subunit chain.
The method of any one of the preceding claims, wherein the antibody is attached to a solid support.
The method according to any one of the preceding claims, wherein the antibody is labeled with one or more fluorophore(s).
The method according to any one of the preceding claims, wherein the fluorophore is phycoerythrin, allophycocyanin, fluorescein, Texas red, or Alexa Fluor 647, brilliant dyes.
The method according to any one of the preceding claims wherein the antibody further comprises a moiety suitable for detection.
The method according to any one of the preceding claims, wherein the moiety suitable for detection is selected from nanoparticles, and radioisotopes.
The method according to any one of the preceding claims, wherein the nanoparticle is selected from the group consisting of micelles, inelastic shells, nanotubular particles, liposomes, gold nanoparticles and polymers.
The method according to any one of the preceding claims, wherein the radioisotope is selected from the group consisting of beta-emitters, auger-emitters, conversion electron-emitters, alpha-emitters, and low photon energy-emitters.
The method according to any one of the preceding claims, wherein the detectable moiety comprises or consists of a paramagnetic isotope.
The method according to any one of the preceding claims, wherein the paramagnetic isotope is selected from the group consisting of 157Gd, 55Mn, 162Dy, 52Cr and 56Fe.
The method according to any one of the preceding claims, wherein the detectable moiety is detectable by an imaging technique such as SPECT, PET, MRI, optical or ultrasound imaging.
A method for manufacturing an isolated population of mammalian cells in vitro which are enriched for neural stem cells and/or neural progenitor cells relative to a reference population, the method comprising the steps of
- a) providing at least a portion of a population of cells, or a portion of a reference population, comprising a neural stem cell and/or a neural progenitor cell,
- b) introducing into the population of cells in a) above a compound identifying an integrin alpha10 subunit expressed by the neural stem cell and/or neural progenitor cell,
- c) selecting and isolating, or isolating and selecting, or isolating, or selecting, the neural stem cells and/or the neural progenitor cells from the population of cells in step b) above,
- thereby producing a population of cells enriched for neural stem cells and/or neural progenitor cells.
An in vitro cell culture of undifferentiated mammalian cells expressing an integrin alpha10 subunit, wherein the cells are derived from neural tissue and wherein
- a) cells in the culture have the capacity to differentiate into neurons and/or oligodendrocytes and/or astrocytes when differentiated in a culture medium substantially free of both serum and a proliferation-inducing growth factor as defined in (b) to produce a cell culture of at least 10% neurons and/or oligodendrocytes and/or astrocytes;
- b) the cell culture divides in a culture medium containing a serum replacement such as B27 and at least one proliferation-inducing growth factor;
- c) cells in the culture differentiate into neurons and/or oligodendrocytes and/or astrocytes upon withdrawal of both the serum replacement and the proliferation inducing growth factor.
An in vitro cell culture comprising
- a) a culture medium containing a serum replacement such as B27 and at least one proliferation-inducing growth factor;
- and
- b) undifferentiated mammalian cells derived from the central nervous system of a mammal, wherein at wherein at least 10%, preferably at least 20%, preferably at least 30%, preferably at least 40%, preferably at least 50%, preferably at least 60%, preferably at least 70%, preferably at least 80%, preferably at least 90% of the cells express an integrin alpha10 subunit.
A suspension culture of mammalian undifferentiated cells expressing an integrin alpha10 subunit, wherein said cells are substantially formed into cell aggregates, and wherein the cell aggregates are maintained in a culture medium containing a proliferation-inducing growth factor.
A method of treating disease or damage and/or preventing and protecting from damage of the nervous system in a subject in need thereof, the method comprising:
- a) providing a composition comprising an enriched population of mammalian neural stem cells and/or mammalian neural progenitor cells, wherein the cells express an integrin alpha10 subunit;
- b) administering a therapeutically effective amount of the isolated population of mammalian neural stem cells and/or neural progenitor cells to the subject,
- thereby treating the disease or damage and/or preventing and protecting from damage of the central nervous system.
A method of treating a mental and behavioral disorder in a subject in need thereof, the method comprising:
- a) providing a composition comprising an enriched population of mammalian neural stem cells and/or mammalian neural progenitor cells, wherein the cells express an integrin alpha10 subunit;
- b) administering a therapeutically effective amount of the isolated population of mammalian neural stem cells and/or neural progenitor cells to the subject,
- thereby treating the neurologic disorders with psychiatric symptoms.
A method of treating disease or damage and/or preventing and protecting from damage of the nervous system in a subject in need thereof, the method comprising:
- a) providing a composition comprising an enriched population of mammalian mesenchymal stem cells, wherein the cells express integrin alpha10 subunit;
- b) administering a therapeutically effective amount of the isolated population of mammalian mesenchymal stem cells to the subject,
- thereby treating the disease or damage and/or preventing and protecting from damage of the central nervous system.
A method of treating a mental or behavioral disorder in a subject in need thereof, the method comprising:
- a) providing a composition comprising an enriched population of mammalian mesenchymal stem cells, wherein the cells express integrin alpha10subunit;
- b) administering a therapeutically effective amount of the isolated population of mammalian mesenchymal stem cells to the subject,
- thereby treating the neurologic disorders with psychiatric symptoms.
An in vitro method for determining the characteristics of a damaged or diseased area of the CNS in a patient in need thereof, the method comprising the steps of:
- a) administering an anti-integrin alpha 10 subunit to a subject,
- b) detecting expression of integrin alpha10 subunit in the damaged or diseased area of the CNS of the subject,
- c) determining characteristics such as location and size of the damaged or diseased area of the CNS.
A marker for mammalian neural stem cells and/or mammalian neural progenitor cells, comprising an integrin alpha10 chain subunit expressed by the neural stem cell and/or neural progenitor cells, for use in a method of treating a disease or damage and/or for preventing and protecting from damage of the nervous system and/or for treating a mental and behavioral disorder, or for diagnosing and/or characterizing the disease or damage.
A composition comprising an isolated population of mammalian neural stem cells and/or mammalian neural progenitor cells expressing an integrin alpha10 subunit for use in a method of treatment of disease or damage of the nervous system and/or mental and behavioral disorder
The method, the marker for use, or the composition according to any one of the preceding claims wherein the neural stem cell is an allogenic neural stem cell.
The method, the marker for use, or the composition according to any one of the preceding claims wherein the neural stem cell is an autologous neural stem cell.
The method, the marker for use, or the composition according to any one of the preceding claims wherein the neural progenitor cell is an allogenic neural progenitor cell.
The method, the marker for use, or the composition according to any one of the preceding claims wherein the neural progenitor cell is an autologous neural progenitor cell.
The method, the marker for use, or the composition according to any one of the preceding claims wherein the mesenchymal stem cell is an allogenic mesenchymal stem cell.
The method, the marker for use, or the composition according to any one of the preceding claims wherein the mesenchymal stem cell is an autologous mesenchymal stem cell.
Dependencies are inferred from the claim text.