The present disclosure relates to methods for generating microglial cells derived from stem cells (e.g., human stem cells), microglial cells obtained from such methods and compositions comprising thereof, and uses of said microglial cells for disease modeling and for treating microglia related disorders.

Disclosed is a method for obtaining a population of human Treg cells including the steps of: (a) culturing a population of human monocytes with a medium including an amount of an interleukin-34 (IL-34) polypeptide in order to obtain a population of immunosuppressive macrophages; (b) co-culturing a population of human peripheral blood mononuclear cells (PBMCs) and the population of immunosuppressive macrophages obtained at step (a).

Disclosed herein are various embodiments relating to methods of producing iMGLs, for example, from pluripoteiit stem cells (PSCs), methods of using iMGLs, and compositions of iMGLs. Also disclosed herein are methods to study various neurological disorders, such as for studying Alzheimer's disease. In addition, disclosed herein are methods of investigating genotypic and phenotypic effects of microglia cells in various physiological and pathological environments in the CNS and brain.

Disclosed is a method for labeling, detecting and/or isolating Foxp3+ Treg cells from a biological sample containing peripheral blood mononuclear cells (PBMC) or lymphocytes including the following steps of: (i) coupling the surface of PBMC or lymphocytes to a capture moiety which binds to the cell through a cell surface molecule and to interleukin-34 (IL34), (ii) culturing the lymphocytes under conditions wherein IL34 is secreted, released and specifically captured by the capture moiety, (iii) labeling the IL34 expressing lymphocytes with a label moiety, and (iv) optionally detecting and/or isolating the IL34 expressing lymphocytes which are Foxp3+ Treg cells. Also disclosed is an isolated population of Foxp3+ Treg cells obtainable by the method and uses thereof.

The purpose of the present invention is to efficiently produce microglia from pluripotent stem cells. Provided is a method for producing microglia from pluripotent stem cells, comprising the following steps: (a) a step of co-culturing a pluripotent stem cell together with a feeder cell for 7 days or longer, and obtaining a blood progenitor cell; (b) a step of co-culturing the blood progenitor cell obtained in step (a) together with a feeder cell in the presence of IL-3 and/or GM-CSF, and obtaining an embryonic monocyte; and (c) a step of, in the presence of M-CSF, co-culturing the embryonic monocyte obtained in step (b) together with an astrocyte, or culturing the embryonic monocyte using an astrocyte supernatant.

The present invention provides a method of inducing microglia cells from blood cells, comprising culturing the blood cells in the presence of interleukin-34 (IL-34) and granulocyte-macrophage colony stimulating factor (GM-CSF).

Provided is a method for producing microglia, including: Step (S1) of inducing differentiation of hemangioblasts to obtain microglial progenitor cells; and Step (S2) of inducing differentiation of the microglial progenitor cells to obtain microglia, in which, in the step of obtaining microglial progenitor cells, expression of PU.1 transcription factor encoded by an exogenous gene is induced, and culture is carried out in the presence of FGF2, SCF, IL-3, IL-6, VEGF, and Wnt inhibitor.

Disclosed is a method for obtaining a population of human Treg cells including the steps of: (a) culturing a population of human monocytes with a medium including an amount of an interleukin-34 (IL-34) polypeptide in order to obtain a population of immunosuppressive macrophages; (b) co-culturing a population of human peripheral blood mononuclear cells (PBMCs) and the population of immunosuppressive macrophages obtained at step (a).

Mononuclear phagocytes derived from induced pluripotent stem cells, denoted as iMPs, which comprise monocytes generated from the induced pluripotent stem cells and optionally further macrophages generated from the induced pluripotent stem cells, are provided for use in improving cognitive function, improving neural health, and/or alleviating or treating a neurodegenerative disorder in a mammal In various embodiments, the iMPs produce macrophages after transplantation or after being stimulated in vitro, and/or express macrophage markers. We showed that iMPs upon administration improve cognition and neural healthy in rodent models of aging, Alzheimer's disease, and amyotrophic lateral sclerosis. Treatment methods are also provided using mononuclear phagocytes generated from autologous stem cells or from induced pluripotent stem cells obtained from autologous cells in patients in need of treatment or prophylaxis of a neurodegenerative disorder.

Disclosed is a method for labeling, detecting and/or isolating Foxp3+ Treg cells from a biological sample containing peripheral blood mononuclear cells (PBMC) or lymphocytes including the following steps of: (i) coupling the surface of PBMC or lymphocytes to a capture moiety which binds to the cell through a cell surface molecule and to interleukin-34 (IL34), (ii) culturing the lymphocytes under conditions wherein IL34 is secreted, released and specifically captured by the capture moiety, (iii) labeling the IL34 expressing lymphocytes with a label moiety, and (iv) optionally detecting and/or isolating the IL34 expressing lymphocytes which are Foxp3+ Treg cells. Also disclosed is an isolated population of Foxp3+ Treg cells obtainable by the method and uses thereof.