Cocktails of chemical inducers of neuron-like properties (CINP) is provided, which includes cAMP agonists, neurogenic small molecules, glycogen synthase kinase inhibitors, TGF receptor inhibitors, and BET family bromodomain inhibitors and optionally, a selective inhibitor of ROCK or p38 MAPK. These cocktails are used in a method of inducing neuron-like properties in partially or completely differentiated non-neuronal cells. The method includes contacting cells of a first type (non-neuronal) with the CINPs for a sufficient period of time to result in reprogramming the cell into cells of a second type having neuron-like characteristics (CiNs). Isolated chemically induced neurons (CiNs) can be used in a number of applications, including but not limited to cell therapy.

Described herein are cell culture media useful for the differentiation of human pluripotent stem cells into microglia. The methods described herein relate to in vitro generation of expandable, bankable, microglial cells by directed differentiation from human pluripotent stem cells (induced or embryonic). Using only defined cell culture media, differentiation of pluripotent stem cells is directed down a mesodermal path, in a rapid and scalable fashion, to generate cells adopting signatures of their in vivo counterparts, including gene expression, protein marker expression and functionality.

The invention provides a cell-containing vessel comprising a plurality of neural cell-containing spheroids for which the variation calculated using the calculation method described below is less than 20%, wherein the neural cell-containing spheroids contain a plurality of types of cells including neural cells,

Calculation Method

Calcium transient assays are conducted for each of a plurality of neural cell-containing spheroids, a number of spontaneous oscillations in a 10-minute period is measured, an average and standard deviation for the number of spontaneous oscillations are calculated, and a variation is calculated using formula (1) below:

[00001]$\begin{array}{cc}\mathrm{Variation}\left(\%\right)=\mathrm{standard}\mathrm{deviation}\mathrm{for}\mathrm{number}\mathrm{of}\mathrm{spontaneous}\mathrm{oscillations}/\mathrm{average}\mathrm{number}\mathrm{of}\mathrm{spontaneous}\mathrm{oscillations}100.& \left(1\right)\end{array}$

The invention relates to an in vitro cell culture comprising neuronal cells derived from iPSC, optionally with an NGN2 transgene, astrocytes derived from iPSC with a SOX9 transgene, and oligodendrocytes derived from iPSC with a SOX10 transgene.

The present disclosure provides a method of directly converting a stem cell into a lineage specific cell, comprising the steps of a) transfecting a stem cell with at least one expression vector comprising i) one or more cell lineage reprogramming factors operably linked to an inducible promoter and ii) a selection marker; and b) inducing the transfected stem cell from stem a) with an inducing agent to directly convert said stem cell into a lineage-specific cell. Particularly exemplified are methods of transfecting a stem cell with SA-ASCL1 (phospho-mutant), DLX2, LHX6 and miR-9/9*-124 linked to a doxycycline inducible promoter to convert the stem cell into an inhibitory neuron and transfecting with NeuroD2 linked to a doxycycline inducible promoter to convert a stem cell into an excitatory neuron. Methods of screening one or more factors and/or one or more genetic mutations that modulate a pre-selected activity of the lineage specific cell, kits and directly convertible stem cells obtained using method of the invention are also provided.

Synthetic human blood vessels can be constructed using human brain derived endothelial cells and incorporated into a tissue model that contains astrocytes and other neurons and microglia. Multi-cell type microvessels incorporate cell types such as astrocytes and pericytes in order to construct a highly representative blood-brain barrier in vitro model with a functional lumen containing brain-derived microvascular endothelial cells and a polymer wall containing human astrocytes and/or pericytes.

The present disclosure generally relates to a process to prepare a cell culture system that mimics the structure of blood brain barrier (BBB) and are useful to study the functions thereof. In particular, the present invention relates to a direct-contact coculture and triculture systems prepared by plating BMECs on a pre-formed lawn of coculture of astrocytes and pericytes on the apical surface of a culture-chamber to achieve a truly direct contact triculture model for BBB. The cell culture systems disclosed herein are also useful for studying the functions of the blood brain barrier and predicting the efficacy and potential toxicity of a drug candidate.

A model blood brain barrier obtained from hPSCs is disclosed.

This disclosure relates generally to membrane-bound compositions, in particular exophers having a diameter between 1 and 20 microns induced from human cells, and uses thereof.

The present invention relates to a method for producing biocompatible structures for the connection and cultivation of biological material, a method for cultivating aggregates of biological material, and the use of biocompatible structures for the connection and cultivation of biological material.