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 present disclosure belongs to the field of biological medicines, and relates to an inducer for inducing differentiation of mesenchymal stem cells into estradiol-secreting cells. The inducer for inducing differentiation of mesenchymal stem cells into estradiol-secreting cells uses human mesenchymal stem cell serum-free culture medium as a substrate and comprises the following components in mass concentration ratios: 20-60 mg/L of bone morphogenetic protein-4, 20-60 mg/L of bone morphogenetic protein-7, 2-8 mg/L of retinoic acid, 2-8 mg/L of resveratrol, 2-8 mg/L of icariin, 2-8 g/L of benzamide, 2-8 g/L of chloroplatinic acid hexahydrate, 2-8 g/L of ethanolamine, 2-10 g/L of erythropoietin and 2-10 g/L of vascular endothelial growth factor. The inducer for inducing differentiation of mesenchymal stem cells into estradiol-secreting cells provided by the present disclosure has a high induction efficiency.

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 invention provides methods for engrafting a bone marrow organoid with CD34+ hematopoietic stem and progenitor cells. Compositions comprising the factors needed for engraftment are also provided.

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.

The present disclosure relates to a SURF4 gene-knockout erythroid progenitor cell and a method of differentiating the same into an erythrocyte, wherein it has been determined that SURF4 gene-knockout cells, in which a SURF4 gene is knocked out in erythroid progenitor cells, express erythroid differentiation markers at a higher rate and undergo erythroid differentiation more rapidly under erythroid differentiation conditions according to the present disclosure, such that the present disclosure provides a method that enables rapid differentiation of erythrocytes using SURF4 gene-knockout erythroid progenitor cells.

The present disclosure relates to a UT2 gene-knockout erythroid progenitor cell and a method of differentiating the same into an erythrocyte, wherein it has been determined that UT2 gene-knockout cells, in which a UT2 gene is knocked out in erythroid progenitor cells, express erythroid differentiation markers at a higher rate and undergo erythroid differentiation more rapidly under erythroid differentiation conditions according to the present disclosure, such that the present disclosure provides a method that enables rapid differentiation of erythrocytes using UT2 gene-knockout erythroid progenitor cells.

The present disclosure relates to an ERR gene-knockout erythroid progenitor cell and a method of differentiating the same into an erythrocyte, wherein it has been determined that ERR gene-knockout cells, in which an ERR gene is knocked out in erythroid progenitor cells, express erythroid differentiation markers at a higher rate and undergo erythroid differentiation more rapidly under erythroid differentiation conditions according to the present disclosure, such that the present disclosure provides a method that enables rapid differentiation of erythrocytes using ERR gene-knockout erythroid progenitor cells.