Provided are methods and compositions for obtaining functionally enhanced derivative effector cells obtained from directed differentiation of genomically engineered iPSCs. The derivative cells provided herein have stable and functional genome editing that delivers improved or enhanced therapeutic effects. Also provided are therapeutic compositions and the use thereof comprising the functionally enhanced derivative effector cells alone, or with antibodies or checkpoint inhibitors in combination therapies.

Provided are methods of deriving a mammalian livestock pluripotent stem cells line, by (a) ex-vivo culturing a mammalian livestock embryo of at least 7 days post-fertilization for a culturing period of at least 4 days and no more than until 21 days post-fertilization so at to obtain an embryo comprising epiblast cell and/or late stage pluripotent stem cell; (b) isolating from the embryo the epiblast cell and/or the late stage pluripotent stem cell, and (c) culturing the epiblast cell and/or the late-stage pluripotent stem cell under conditions suitable for expansion of undifferentiated mammalian livestock pluripotent stem cells to thereby obtain a population of mammalian livestock pluripotent stem cells. Also provided are isolated mammalian livestock pluripotent stem cells, and cells differentiated therefrom.

Methods for cryopreservation of biological samples are provided. The biological samples are sub-millimeter or millimeter scale biological materials. The biological samples are pancreatic islets and stem cell derived islets. Methods for cryopreservation of islets using cryomesh and multi-step loading and unloading of CPA cocktails are provided. Methods disclosed result in vitrified and rewarmed islets with high recovery, viability and functionality. Methods are scalable for high throughput production of large amounts of vitrified and rewarmed islets for use in therapeutic transplantation.

The present disclosure provides methods for bulk droplet vitrification of cells, compositions including the vitrified droplets, and systems for performing the methods for bulk droplet vitrification cells.

The present application relates to the field of cell biology. The present application provides a cryopreservation medium for cryogenic storage of a biological sample, particularly a cryopreservation medium for cryogenic storage of cells in an apheresis sample.

The purpose of the present invention is to obtain a cell suspension with a low concentration of cryoprotectant. This method of preserving cells used comprises the steps of (a) enriching cells from a cell suspension containing the cells and a cryoprotective solution to generate an enriched fraction, and (b) freezing the enriched fraction to prepare a frozen material. It is also possible to use a method of producing a cell suspension, comprising the steps of (a) enriching cells from a cell suspension containing the cells and a cryoprotective solution to generate an enriched fraction, (b) freezing the enriched fraction to prepare a frozen material, (c) thawing the frozen material to prepare a thawed material, and (d) mixing the thawed material and a solution to produce a cell suspension.

A population of enteroendocrine cells (EEC) is obtained from a mammalian post-natal cell population, such as a population including post-natal stem cells, by treating the population with a plurality of small molecules that upregulate ChgA and promote differentiation of the cells to form the enteroendocrine cells. The upregulation of ChgA is such that the fraction of cells expressing CGA in the obtained cell population, as measured by a ChgA Immunostaining Assay, is at least about 1.5%. Small molecules that can be used to differentiate the post-natal cells into the enteroendocrine cells can include at least one of a Wnt activator, a Notch inhibitor, a Wnt inhibitor, a MEK/ERK inhibitor, a growth factor, a HDAC inhibitor, a Histone Methylation Inhibitor, a Tgf-β inhibitor, and a NeuroD1 activator. Also, the insulin expression of a population of mammalian cells is increased by treating the population with a plurality of small molecules that increase the insulin expression.

Methods for generating serum-free and/or xenogen-free cardiac explant-derived stem cells (EDC) are provided. These methods may include providing an initial cardiac explant, which has been minced and digested; plating the initial cardiac explant; culturing the plated cardic explant in serum-free and xenogen-free medium; harvesting EDC cells surrounding or emerging from the plated cardiac explant; and optionally performing static expansion of harvested EDC cells in serum-free and xenogen-free media. Serum-free and/or xenogen-free cardiac EDC cells produced by these methods, as well as methods and uses thereof for the treatment of heart failure in a subject in need thereof, are also provided.

This invention provides a system for efficiently producing differentiated cells from pluripotent cells, such as human embryonic stem cells. Rather than permitting the cells to form embryoid bodies according to established techniques, differentiation is effected directly in monolayer culture on a suitable solid surface. The cells are either plated directly onto a differentiation-promoting surface, or grown initially on the solid surface in the absence of feeder cells and then exchanged into a medium that assists in the differentiation process. The solid surface and the culture medium can be chosen to direct differentiation down a particular pathway, generating a cell population that is remarkably uniform. The methodology is well adapted to bulk production of committed precursor and terminally differentiated cells for use in drug screening or regenerative medicine.

Induced pluripotent stem cells are treated using a combination of compounds that improve competence of the induced pluripotent stem cells in responding to differentiation signals and/or improve the efficiency of differentiation of the treated induced pluripotent stem cells in differentiation towards a desired phenotype. The combination treatment can incorporate two or more of prolongation of early G1 phase, treatment with an interleukin, modulation of DNA methylation, modulation of histone acetylation, and activation of the Wnt pathway. Cells derived from induced pluripotent stem cells so treated can be used in regenerative therapy and production of organoids.