The present invention refers to a cell suspension of autologous or allogeneic, preferably autologous, adult bone marrow-derived white blood cells, preferably derived from the crest of the ilium (or iliac crest), enriched for mononuclear cells, meaning more than 25% of the white blood cells (WBCs) present in the cell suspension are mononuclear cells, and comprising: A. Mononuclear cells (MNC) selected from the list comprising or consisting of: i. A population of Lymphocytes; ii. A population of Monocytes; and iii. A population of Hematopoietic stem cells expressing CD34;
and wherein the cell suspension further comprises: B. Granulocytes;
for use in the treatment or amelioration of lower extremity peripheral artery disease.

A blood product containing peripheral blood mononuclear cells (PBMCs) in an amount of at least 4 million cells per milliliter and human chorionic gonadotropin (HCG in an amount of at least 150 international units (IU) per milliliter. A method of preparing the blood product, including applying HCG to a female patient, then obtaining PBMCs from the female patient, then adding HCG to the obtained PBMCs. A method of culturing PBMCs, including applying HCG to a female patient, then culturing PBMCs obtained from the female patient at a time after the HCG was applied to the patient. A method of in vitro fertilization, including applying HCG to a female patient, culturing PBMCs obtained from the patient after the HCG was applied to the patient, introducing the cultured PBMCs into the uterus of the patient, and transferring at least one embryo into the uterus of the patient.

A population of isolated mammalian macrophages, the cell surface of which is chemically modified to comprise a molecule that comprises an imaging agent or a therapeutic agent or a molecule that binds to an imaging agent or a therapeutic agent, and methods of making and using the modified macrophages are provided.

The present invention provides a regulatory T (Treg) cell expressing an antigen chimeric receptor (CAR) comprising a) at least one antigen binding domain, b) a transmembrane domain, and c) a cytoplasmic signaling domain comprising at least one primary cytoplasmic signaling domain and at least the co-stimulatory signaling domain of CD137, wherein said antigen binding domain specifically binds an antigen that is expressed on the surface of a target cell or a tag of a tagged polypeptide that binds to an antigen expressed on the surface of a target cell or a soluble antigen. Compositions comprising said Treg cells, and methods of enrichment and analysis of activated Tregs that express said CAR are also disclosed.

Some embodiments provided herein relate to methods and compositions for making genetically modified T cells. In some such embodiments, CD4+ and CD8+ T cells are cultured in a single serum-free volume. In some embodiments, co-cultured CD4+ and CD8+ T cells can be transduced with a lentiviral vector, and a population of transduced T cells can be harvested within a shorter period of time than other conventional methods.

Provided herein are compositions and methods for modifying T cells. The disclosure is based, in part, on the use of sgRNA lentiviral infection with Cas9 protein electroporation (SLICE), to identify regulators of IL2RA, IL-2, CTLA4, arnd FOXP3 in effector T cells. IL2RA, IL-2, CTLA4, and FOXP3 are key genes in immune regulation that have been implicated in autoimmune disease and cancer. Therefore, modulating expression of these genes in T cells, for example, effector T cells or regulatory T cells, could have therapeutic applications.

Systems and methods for sorting T cells are disclosed. Autofluorescence data is acquired from individual cells. An activation value is computed using one or more autofluorescence endpoints as an input. The one or more autofluorescence endpoints includes NAD(P)H shortest fluorescence lifetime amplitude component (.sub.1).