The present disclosure relates to improved methods serum free stem cell culture, particularly 3D culture in bioreactors as well as cell culture medium and compositions for use in the same. Such methods may be particularly suitable for large scale cell manufacture.

There is provided a cell culture matrix comprising a fungal derived protein. Also provided is a composition comprising the cell culture matrix as described herein, a cell culture system comprising the cell culture matrix as described herein, and a method of forming a cell culture matrix thereof.

The present disclosure provides, in part, a cell culture medium supplement comprising at least one plant protein homologue of a serum protein, a cell culture medium comprising a serum-free base medium and one or more plant based proteins, and methods of growing cells in vitro and of producing cultured meat using the cell culture medium.

Provided are novel serum-free culture media which comprise basic fibroblast growth factor (bFGF), transforming growth factor beta-3 and ascorbic acid at a concentration of at least about 50 microgram/ml; ascorbic acid at a concentration range of about 400-600 microgram/ml, bFGF at a concentration range of about 50-200 ng/ml, xeno-free serum replacement and a lipid mixture; the IL6RIL6 chimera at a concentration range of about 50-200 picogram per milliliter (pg/ml); or leukemia inhibitory factor (LIF) at a concentration of at least 2000 units/ml; cell cultures comprising same with pluripotent stem cells such as human embryonic stem cells and induced pluripotent stem (iPS) cells, and methods of using same for expanding pluripotent stem cells in an undifferentiated state using two-dimensional or three-dimensional culture systems; and methods of expanding iPS cells in a suspension culture devoid of substrate adherence and cell encapsulation.

A method for generating a population of cardiomyocytes from induced pluripotent stem cells (iPSCs) in an integrated process. The method may comprise seeding the iPSCs on a modified surface of a modified cell culture substrate, culturing the seeded iPSCs on the modified surface of the modified cell culture substrate in an animal component-free culture medium, and differentiating the cultured iPSCs to the population of cardiomyocytes on the modified surface of the modified cell culture substrate. The modified cell culture substrate may comprise a patterned polydimethylsiloxane (PDMS) substrate, a first coating comprising a plurality of polydopamine molecules, and a second coating comprising a plurality of Laminin 511 E8 Fragment (LME8) molecules.

Alveolar-like macrophages and a method for generating alveolar-like macrophages from hemangioblasts is provided. The method comprises the steps of: i) culturing the hemangioblasts in a hematopoietic-inducing medium comprising vascular endothelial growth factor (VEGF), stem cell factor (SCF) and interleukin-3 (IL-3) for a sufficient period of time to generate macrophages, and ii) culturing the macrophages in an alveolar macrophage-inducing medium comprising granulocyte macrophage colony stimulating factor (GM-CSF), and optionally macrophage colony stimulating factor (M-CSF), under suitable conditions and for a sufficient period of time to yield alveolar-like macrophages.

Disclosed herein is the cell culturing supernatant from human decidual mesenchymal stem cell culturing that are therapeutic to a subject having an ischemic disease, such as, diabetic foot ulcer. Through the establishment of a mouse hindlimb ischemia model, treatment with cell culturing supernatant from human decidual mesenchymal stem cell culturing can improve the blood supply of the hind limbs. In the streptozotocin induced diabetic model, treatment with cell culturing supernatant from human decidual mesenchymal stem cell culturing can also improve the blood supply of the hind limbs. The invention can be applied to diabetic patients to reduce the incidence of amputation by promoting angiogenesis.

Provided is an isolated population of human pluripotent stem cells comprising at least 50% human pluripotent stem cells characterized by an OCT4+/TRA1-60−/TRA1-81−/SSEA1+/SSEA4− expression signature, and novel methods of generating and maintaining same in a pluripotent, undifferentiated state a suspension culture devoid of cell clumps. Also provided are novel culture media, cell cultures and methods for culturing pluripotent stem cells in a suspension culture or a two-dimensional culture system while maintaining the cells in a proliferative, pluripotent and undifferentiated state. The novel culture media comprise interleukin 11 (IL11) and Ciliary Neurotrophic Factor (CNTF); bFGF at a concentration of at least 50 ng/ml and an IL6RIL6 chimera; or an animal contaminant-free serum replacement and an IL6RIL6 chimera. Also provided are methods for generating lineage-specific cells from the pluripotent stem cells.

Provided herein are methods and compositions related to culturing hemoglobin-dependent bacteria.

Methods for generating high-yield, high-purity cardiomyocyte progenitors or cardiomyocytes from pluripotent cells are described. Wnt/β-catenin signaling is first activated in pluripotent cells, by, for example, inhibiting Gsk-3 to obtain a first population of cells. Wnt/β-catenin signaling is then inhibited in the first cell population to induce cardiogenesis. One or more of these steps is performed under defined, albumin-free culture conditions.