The technology described herein is directed to improved methods of T cell differentiation. Also described herein are immune cells differentiated using such methods and compositions comprising such immune cells. In some embodiments, the immune cells can be genetically modified. In some embodiments, the immune cells or compositions comprising said immune cells can be administered to a patient as a cellular replacement therapy to treat a condition.

In vitro process for producing organoids from mammalian cells, the process comprising or the steps of cultivating pluripotent stem cells (PSC) in cell culture medium in a stirred tank bioreactor under conditions suitable for producing PSC aggregates, removing PSC aggregates suspended in cell culture medium from the bioreactor, and preferably in a flow channel, encapsulating PSC aggregates separately in biocompatible hydrogel to produce separate hydrogel-encapsulated PSC aggregates, incubating the hydro-gel-encapsulated PSC aggregates in cultivation vessels under static conditions in medium containing at least one differentiation factor.

The disclosure provides methods of making CAR-expressing immune effector cells (e.g., T cells, or NK cells), and compositions and reaction mixtures comprising the same. The disclosure further provides methods of using said CAR-expressing immune effector cells.

The invention provides culture platforms, cell media, and methods of differentiating pluripotent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein, which enable feed-free, monolayer culturing and in the absence of EB formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, NK cells, NKT cells and B cells.

The invention provides culture platforms, cell media, and methods of differentiating pluripotent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein, which enable feed-free, monolayer culturing and in the absence of EB formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, and NK cells.

The present disclosure, in various aspects and embodiments, provides methods for generating hematopoietic lineages for cell therapy, including erythroid progenitor cells, progenitor erythroblasts, granulocyte-macrophage progenitor cells (GMPs), and megakaryocyte erythroid progenitor cells (MEPs), and erythroid cells. In various embodiments, the invention provides for efficient ex vivo processes for developing such hematopoietic lineages, including but not limited to progenitor erythroblast cells and erythroblast cell lineages, from human induced pluripotent stem cells (iPSCs). Cells generated according to the disclosure in various embodiments are functional and/or more closely resemble the corresponding lineage isolated from peripheral blood or bone marrow. The present invention also provides isolated cells and cell compositions produced by the methods disclosed herein, as well as methods for cell therapy.