The present disclosure provides compositions and methods comprising human platelet releasate (hPR), a xeno-free media supplement. The disclosure also relates to a cGMP process for the rapid, efficient and large-scale manufacturing of hPR that may be performed in less than 4 hours. The platelet releasate of the current disclosure prevents gelation of growth media alleviating the need for heparin or other anticoagulants. Mesenchymal stem cells expanded in the presence of platelet releasate have demonstrated superior expansion rates and potency compared to commercial supplements including platelet lysates. The releasate has therapeutic and medical applications. The disclosure also relates to compositions and methods related to platelet-rich fibrin.

The present invention provides a method for preparing 3D brain organoids, comprising the following steps: neurospheres obtained by the RONA method are dissociated into single cells by accutase, plated on a cell culture plate after being counted, cultured in medium A until day 7; neurospheres are cultured in medium B until day 25˜35, and then they are encapsulated by Matrigel; neurospheres are further cultured in media B until day 55˜65, and then they are encapsulated by Matrigel for the second time and cultured continually afterwards. The present invention also provides a medium for culturing 3D brain organoids. The present invention begins with highly purified neurospheres obtained by the RONA method, and neuronal stem cells can be controlled and cultured to achieve true 3D brain organoids with uniform size and structure by this relatively simple method. The 3D brain organoids have six-layered cortical structure of the brain and various subtypes of inhibitory interneuron cells, which are suitable for disease research in vitro, drug screening, etc., and are of great significance in industrialization.

A bioreactor system for conditioning of pluripotent cells or cell media is provided. In further aspects, conditioned pluripotent cells and methods for making such cells are provided.

Efficient stem cell delivery into biomaterials using capillary driven encapsulation are disclosed herein where stem/progenitor and/or tissue specific cells are rapidly and efficiently seeded via capillary driven encapsulation into a porous scaffold for cell delivery in the skin or any other organ. The rapid capillary force approach maximizes both seeding time and efficiency by combining hydrophobic, entropic and capillary forces to promote active, ‘bottom-up’ cell engraftment. This methodology uses micro domain patterned biopolymers in a porous dry gel to generate capillary pressure to move a viscous stem cell mix from a hydrophobic reservoir into the polymer matrix to promote active cell seeding within the entire gel volume.

The subject invention concerns materials and methods for expansion of stem cells, such as mesenchymal stem cells (MSC), that improve translational success of the cells in the treatment of various conditions. The subject invention utilizes cell self-aggregation as a non-genetic means to enhance their therapeutic potency in a microcarrier bioreactor. In one embodiment of the method cells are cultured in a container or vessel in the presence of thermally responsive microcarriers (TRMs) wherein cells adhere to the surface of the TRMs. After a period of time the cell culture temperature is reduced so that the cells detach from the TRMs. The detached cells are allowed to form 3D aggregates. The 3D aggregates can be collected and treated to dissociate the cells. Dissociated cells can then be used for transplantation in methods of treatment or for in vitro characterization and study.

The present invention relates to a method for in vitro production of red blood cells. The method for in vitro production of red blood cells, according to the present invention, specifies, by cell size, the maturation step of cells exhibiting optimal effects in an agitation-type culture, so that even if an expert does not always identify cell shape, automated processes of culturing are possible, and thus automation in bioreactors is possible in the mass-production of uniform quality and red blood cells.

The present invention relates to a medium for direct differentiation of embryonic stem cell-derived mesenchymal stem cells, a method of preparing mesenchymal stem cells by using same, mesenchymal stem cells prepared thereby, and a cell therapy product comprising the same mesenchymal stem cells. In a medium composition and a method according to an embodiment, mesenchymal stem cells may be prepared at high yield within a short period of time. In addition, the method is simple in preparation procedure because of the absence of an embryoid body formation step and allows homogeneous cells to be prepared, thus advantageously providing a cell therapy product within a reduce period of time, compared to conventional methods.

A simple, highly flexible and scalable platform for making functional complex tissues with heterogeneity and irregularity is provided. The method includes combining undifferentiated cells, such as pluripotent or multipotent stem cells, with a biomaterial to make multiple undifferentiated or naïve subunits, exposing the undifferentiated or naïve subunits to different cell culture environments for induction of differentiation towards different lineages as required by that complex tissue, and combining the then functional subunits with or without the undifferentiated subunits. The differentiated subunits thus combined can be cultured under biological, chemical, and/or physical culture conditions suitable to fine-tune the structural and functional properties of the bioengineered complex tissue to form a bioengineered tissue graft that mimics the structural and functional characteristics of native complex tissue. The bioengineered tissue graft can then used to replace dysfunctional tissue.

A novel macroscale, contactless, label-free method to print in situ three-dimensional (3D) particle assemblies of different morphologies and sizes is demonstrated using non-adherent (blood) and adherent (MCF-7 and HUVEC) cells. This method of manipulating particles such as cells or biological moleules does not necessarily require the use of nozzles that can contaminate the cell suspension, or to which cells can adhere. Instead, the intrinsic diamagnetic properties of particles such as cells are used to magnetically manipulate them in situ in a nontoxic paramagnetic medium, creating various shapes such as (a) rectangular bar, (b) three-pointed star, and (c) spheroids of varying sizes. A normal distribution of 3D cell structures is produced when formed through magnetic assembly. The use of this method in co-culturing of different cell lines is also demonstrated. The technique is envisioned to be transferable to other cell lines or diamagnetic biological molecules, with potential applications in tissue engineering, medical diagnostics and drug screening.

The present invention relates to providing artificial tendon or ligament tissue having sufficient strength. More specifically, artificial tendon or ligament tissue having sufficient strength is provided by embedding collagen-secreting cells in a gel having strength capable of resisting a tensile load and by culturing the cells while applying a tensile load to the gel to produce artificial tendon or ligament tissue. Cells that steadily express the Mkx gene can be used as the collagen-secreting cells. A fibrin gel containing aprotinin can be used as the gel.