This invention relates to methods and compositions for detecting, identifying and treating glaucoma diseases. More particularly, this invention discloses compositions and methods for affecting intraocular pressure and increasing ocular outflows in glaucoma. Compositions and methods provided include purified and synthesized extracellular vesicle complexes from glaucoma ocular humor for use in methods and devices for detecting, characterizing and treating glaucoma diseases along with active agents. The presence of extracellular vesicle complexes in glaucoma can also be used as a biomarker.

Methods, systems, and compositions are provided for extracting bone marrow cells from bone obtained from deceased donors, for preparing the bone marrow for cryopreservation, and for obtaining desired cells from cryopreserved and fresh bone marrow.

Described herein are in vitro quantitative methods directed to 3D extracellular matrix (ECM) organization and mechanics. The methods do not require application of an external force, anchorage, or a scaffolding material in order to culture ECM tissue constructs, and the methods enable quantitative measurements without crosstalk noise from variables attributable to external forces, anchorage, and scaffolding material.

A mesenchymal stem cell harvesting system and method for increasing the efficiency of collecting and processing physiological fluids containing mesenchymal stem cells from a cavity within a patient's skeletal system. Microenvironments risk in MSC production and concentration within a cavity, for example the patient's ilium, are penetrated with a pointed instrument used to create an aperture in the hard cortical bone forming the cavity followed by the insertion of an aspiration device which extracts one or more samples of cancellous bone, bone marrow, bone marrow blood and other aspirated material. The aspirate is rinsed and may be filtered to remove unwanted material and to increase the concentration and purity of the mesenchymal stem cells in the aspirant far beyond levels formerly obtainable for use in autologous treatment of the patient.

A three-dimensional cell growth medium is described. The cell growth medium may comprise hydrogel particles swollen with a liquid cell growth medium to form a granular gel yield stress material which undergoes a phase transformation from a solid phase to a liquid-like phase when an applied stress exceeds the yield stress. Cells may be placed in the three-dimensional cell growth medium according to any shape or geometry, and may remain in place within the three-dimensional cell growth medium.

The disclosure features methods and compositions for differentiating stem cells into hematopoietic stem and progenitor cells (HSPC) and/or Natural Killer (NK) cells. The methods and compositions described herein are used to differentiate stem or progenitor cells having at least one gene-edit that is maintained in the differentiated cell. Also provided are differentiated cells produced using the methods and compositions described herein for therapeutic applications.

A bioactive suspension derived from freshly disaggregated tissue is provided, as well as related methods of formulation and use. The bioactive suspension may comprise a cell-free supernate derived from epidermal and dermal tissue that has been enzymatically and mechanically disaggregated, then separated, and which may contain tissue regeneration factors known to speed healing. The bioactive suspension may further comprise genetically-modified treatment cells, wild type cells, or both, and may be combined with one or more scaffolding elements to form a bioactive suspension combination product suitable for treatment of a cutaneous defect. Synthetic bioactive suspensions and bioactive suspension combination products are also provided.

A bioactive suspension derived from freshly disaggregated tissue is provided, as well as related methods of formulation and use. The bioactive suspension may comprise a cell-free supernate derived from epidermal and dermal tissue that has been enzymatically and mechanically disaggregated, then separated, and which may contain tissue regeneration factors known to speed healing. The bioactive suspension may further comprise genetically-modified treatment cells, wild type cells, or both, and may be combined with one or more scaffolding elements to form a bioactive suspension combination product suitable for treatment of a cutaneous defect. Synthetic bioactive suspensions and bioactive suspension combination products are also provided.

In some aspects and embodiments, the invention provides methods for making hematopoietic stem cells, including for HSCT. The method comprises providing a cell population comprising hemogenic endothelial (HE) or endothelial cells, and increasing activity or expression of DNA (cytosine-5-)-methyltransferase 3 beta (Dnmt3b) and/or GTPase IMAP Family Member 6 (Gimap6) in the HE and/or endothelial cells under conditions sufficient for stimulating formation of HSCs.

The described invention provides an ex vivo dynamic multiple myeloma cancer niche contained in a pumpless perfusion culture device. The dynamic multiple myeloma cancer niche includes (a) a three-dimensional tissue construct containing a dynamic ex vivo bone marrow niche, which contains a mineralized bone-like tissue containing viable osteoblasts self-organized into cohesive multiple cell layers and an extracellular matrix secreted by the viable adherent osteoblasts; and a microenvironment dynamically perfused by nutrients and dissolved gas molecules; and (b) human myeloma cells seeded from a biospecimen composition comprising mononuclear cells and the multiple myeloma cells. The human myeloma cells are in contact with osteoblasts of the bone marrow niche, and the viability of the human myeloma cells is maintained by the multiple myeloma cancer niche.