The invention provides a defined low protein culture medium for maintaining cells in an undifferentiated state, the medium comprising: a basal medium, an organic acid from the tricarboxylic acid cycle, nonessential amino acids, a combination of growth factors selected from the group consisting of FGF-2 protein, an IGF-1 protein or insulin, a Transferrin protein, and a TGF beta 1 protein, wherein the medium is essentially feeder-free, essentially xeno-free, essentially free of beta-mercaptoethanol, and essentially free of animal-derived or human-derived proteins.

The present disclosure is related to methods for forming a stem cell bank. The methods include obtaining a first stem cell from a multi-cell fertilized embryo, expanding the first stem cell into two or more descendant stem cells, and storing at least one of the descendant stem cells to form the stem cell bank. Also disclosed is a kit that can be used for making the stem cell bank during in vitro fertilization. If desired, the HLA serotype of the stem cells can be determined prior to storage.

The present disclosure relates to a composition for increasing the biological activity of stem cells using mixture 4F. Stem cells treated with mixture 4F according to the present disclosure not only acquire undifferentiated characteristics (stemness), but also have the advantage of improving cell proliferative ability and mobility, and thus, after being transplanted into a body, the stem cells can improve cell survival and engraftment and further enhance the ability to regenerate blood vessels and tissues. Accordingly, the stem cells can have various applications in the fields of stem cell differentiation and ischemic disease prevention or treatment.

A method of generating retinal pigment epithelium cells is disclosed. Cell populations comprising same and uses thereof are also disclosed.

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.

A method for producing eyefield progenitor cells, including: (a) obtaining a starting population comprising human pluripotent stem cells (hPSCs) that are dissociated to essentially single cells; (b) culturing said hPSCs to a contact-inhibited monolayer; (c) contacting said hPSC monolayer in a primary differentiation medium to generate a homogeneous, contact-inhibited monolayer of anterior neuroectodermal cells (ANECs); (d) dissociating said homogeneous ANECs from (c) into essentially single cells; (e) forming dissociated ANECs into size-controlled and homogeneous 3D aggregates (ANEBs), wherein the ANEBs are 3D aggregates of anterior neuroectodermal cells that are distinct from embryoid bodies; and (f) culturing said ANEBs in a primary differentiation medium in suspension to further differentiate them to Eyefield Progenitor Cells (EFPCs).

Mesenchymal stem cells may be efficiently obtained from a biological cell sample containing mesenchymal stem cells by: (1) culturing the biological cell sample containing mesenchymal stem cells in a serum-free medium in the presence of vitronectin or a partial peptide thereof capable of adhering mesenchymal stem cells, and (2) collecting a cell aggregate of the mesenchymal stem cells.

A method for making an acellular human amnion-derived composition configured for therapeutic use is disclosed and generally includes the steps: obtaining amniotic membrane tissue; testing the amniotic membrane tissue for pathogens; washing the amniotic membrane tissue; manually removing blood-containing chorion tissue from the amniotic membrane tissue decellularizing the amniotic membrane tissue with xeno-free enzymes; collecting amniotic cells from the decellularized amniotic membrane tissue; seeding the amniotic cells for culture into xeno-free media formulated for mesenchymal stem cells; growing the amniotic cells to a specified confluency; collecting conditioned media; and freezing the collected conditioned media; wherein the method further includes irradiating the conditioned media.

The current invention relates to a process for the manufacturing of a composition of extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs), said method comprises: culturing and expanding MSCs in a serum-free and xeno-free medium comprising purified human serum albumin and human transferrin; —collecting cell supernatant, said cell supernatant comprises EVs; —filtering said cell supernatant to obtain EVs; and concentrating said EVs, preferably by means of ultrafiltration. In a second and further aspect, said invention is directed to compositions comprising EVs and their clinical use.

The present disclosure provides a process for obtaining an expanded primed mesenchymal stem cell population. In the process, the MSCs are cultured in the culture medium comprising a corneal stromal stem cell derived-conditioned medium to obtain the expanded population of the primed mesenchymal stem cell population along with the mesenchymal stem cell derived-conditioned medium. Also, provided is a method of culturing the MSCs in 3D culture using a spheroid-based method or a microcarrier-based method, in order to obtain the expanded primed mesenchymal stem cell population. Further, an exosome preparation obtained from the expanded primed mesenchymal stem cell derived-conditioned medium is also disclosed herein. The present disclosure also discloses a composition comprising an expanded population of the primed mesenchymal stem cells, or a primed mesenchymal stem cell derived-conditioned medium, or an exosome preparation, or combinations thereof.