Methods for the in vitro production of enucleated red blood cells and the enucleated red blood cells thus prepared are provided. Such enucleated red blood cells may express a sortaggable surface protein, which allows for surface modification in the presence of a sortase. Also described herein are surface modified enucleated red blood cells, e.g., conjugated with an agent of interest such as a peptide, a detectable label, or a chemotherapeutic agent, and uses thereof in delivering the agent to a subject.

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105, CD90, CD66A, CD66E, CXCR4, CD133 or an SSEA. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1, TWIST, KLF-4 and Stella but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105, CD90, CD66A, CD66E, CXCF4, CD133 or an SSEA. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiating the stem cells, are also provided.

Provided herein are methods of isolation and identification of post-natal hemogenic endothelial cells. Further provided are substantially purified populations of post-natal hemogenic endothelial cells, compositions of post-natal hemogenic endothelial cells, and methods to utilize hemogenic endothelial cells to regenerate the hematopoietic system in a subject.

The present invention relates to methods, kits and compositions for expansion of embryonic hematopoietic stem cells and providing hematopoietic function to human patients in need thereof. In one aspect, it relates to kits and compositions comprising a Notch agonist, one or more growth factors, and, optionally, an inhibitor of the TGF pathway. Also provided herein are methods for expanding embryonic hematopoietic stem cells using kits and compositions comprising a Notch agonist, one or more growth factors, and, optionally, an inhibitor of the TGF pathway. The embryonic hematopoietic stem cells expanded using the disclosed kits, compositions and methods include cells derived from an embryo (e.g., aorta-gonad-mesonephros region of the embryo), embryonic stem cells, induced pluripotent stem cells, or reprogrammed cells of other types. The present invention also relates to administering the embryonic hematopoietic stem cells expanded using a combination of a Notch agonist, one or more growth factors, and, optionally, an inhibitor of the TGF pathway to a patient for short-term and/or long-term in vivo repopulation benefits.

The present invention relates to a method for preparing lymphoid progenitors. The inventors took advantage of their original and relevant Warts, Hypogammaglobulinemia, Infections and Myelokathexis (WHIM) Syndrome (WS) model and the access to blood samples from five WS patients to investigate the impact of CXCR4 desensitization on BM and extra-medullary (i.e. splenic) hematopoiesis and hematopoietic stem and progenitor cells (HSPCs) recirculation. They developed, for the first time, an original in vitro system permitting to selectively expand HSPCs to obtain lymphoid progenitors by using an original cocktail of cytokines. In particular, the present invention relates to an in vitro method for preparing lymphoid progenitors by culturing HSPCs in an appropriate culture medium comprising an effective amount of a cocktail of cytokines consisting in SCF, IL-3, IL-6, IL-7, Flt-3, and CXCL12.

Systems and methods to expand hematopoietic stem cell (HSC) using zwitterionic hydrogels (ZTG) are described. Expansion using the disclosed systems and methods results in HSC populations with (i) an increased proportion of HSC versus partially or fully differentiated cells, (ii) proportionally lower cell surface expression levels of differentiation/maturation markers, (iii) reduced metabolic rates following expansion, and/or (iv) a greater proportion of quiescent cells following expansion, as compared to currently available clinical expansion methods.

The present invention relates in part to nucleic acids encoding proteins, nucleic acids containing non-canonical nucleotides, therapeutics comprising nucleic acids, methods, kits, and devices for inducing cells to express proteins, methods, kits, and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, and therapeutics produced using these methods, kits, and devices. Methods for inducing cells to express proteins and for reprogramming and gene-editing cells using RNA are disclosed. Methods for producing cells from patient samples, cells produced using these methods, and therapeutics comprising cells produced using these methods are also disclosed.

Provided is a ??T cell for securing the purity and number of cells sufficient for treatment. Also provided is a method of generating the ??T cell. More specifically, provided are homogeneous ??T cells excellent in that the ??T cells are not affected by exhaustion of the cells. The foregoing is achieved by ??T cells obtained by subjecting induced pluripotent stem cells (iPS cells) to differentiation induction treatment. Specifically, the foregoing is achieved by ??T cells generated by subjecting iPS cells having a rearranged ??TCR gene (??TCR-type iPS cells) to differentiation induction treatment. According to the method of generating the ??T cell of the present invention, there can be provided ??T cells and a cell population of ??T cells that have an excellent function of having antigen-specific cytotoxic activity in a MHC-unrestricted manner, and that are more homogeneous and have a higher effect than ??T cells separated from peripheral blood.

The present specification provides libraries of HLA homozygous induced pluripotent cell

Provided are a method for preparing genetically engineered erythrocytes carrying an anti-PD-1 single chain antibody, and genetically engineered erythrocytes carrying the anti-PD-1 single chain antibody. The preparation method comprises the following steps: constructing a desired fragment sequence in a lentiviral expression vector, lentivirally packaging the vector of the desired sequence to obtain a high-titer lentiviral concentrate; isolating Lin.sup.?CD34.sup.? cells from peripheral blood mononuclear cells and enriching the Lin.sup.?CD34.sup.? cells; inducing the Lin.sup.?CD34.sup.? cells to differentiate into erythroid and performing proliferation thereon; using the lentiviral concentrate to infect the Lin.sup.?CD34.sup.? cells; and obtaining mature anti-PD-1 scFv erythrocytes via erythrocyte denucleation. The present genetically engineered erythrocytes carrying the anti-PD-1 single chain antibody can perform the targeted delivery of an anti-PD-1 single-chain antibody to tumor tissues.