The present disclosure provides the sequence of a Paenibacillus polymyxa preproenzyme which is the precursor of a neutral protease, expression thereof in a transformed host organism, and methods for production of the neutral protease, by recombinant means. Further, use of the recombinantly produced neutral protease is disclosed in the field of cell biology, particularly for the purpose of tissue dissociation. The disclosure also includes blends with other proteases. Further disclosed are nucleotide sequences encoding the neutral protease.

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.

The present disclosure relates to an eyedrop applicable to limbal stem cell deficiency and a preparation method, wherein each liter of the eyedrop includes the following components: 1 to 40 mg of human adipose-derived stem cell exosomes, excipients: 0.5 to 2 g of sodium hyaluronate, 0.5 to 3 g of vitamin B6 and 0.05 to 0.3 g of benzalkonium chloride, and the balance of medical normal saline, a pH of the eyedrop is 6.5 to 7.5; and the eyedrop of the present disclosure is effective in treating limbal stem cell deficiency.

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 method relates to an in vitro method of producing a (three dimensional) neural tissue composition, the method comprising the steps of re-suspending cells that are obtained by culturing pluripotent stem cells in a neural induction medium in cell culture substrate and culturing said resuspended cells in the presence of an neural differentiation medium.

The disclosure provides a chondrocyte cell sheet comprising one or more layers of confluent cells comprising chondrocytes and chondroprogenitor cells. Methods of generating cartilage tissue in a subject are also provided. The disclosure also provides a method for producing chondrocyte cell sheets comprising culturing chondrocytes and chondroprogenitor cells in culture solution on a temperature-responsive polymer which has been coated onto a substrate surface of a cell culture support, wherein the temperature-responsive polymer has a lower critical solution temperature in water of 0-80° C.; adjusting the temperature of the culture solution to below the lower critical solution temperature, whereby the substrate surface is made hydrophilic and adhesion of the cell sheet to the surface is weakened; and detaching the cell sheet from the culture support.

The present invention relates to methods for obtaining skeletal muscle derived cells (SMDC), and the use of SMDCs in a method of preventing and/or treating neuromyopathies and/or myopathies, wherein the neuromyopathy and/or myopathy is incontinence, in particular a urinary and/or an anal or fecal incontinence.

Disclosed herein are compositions and methods for treating, ameliorating or preventing a retinal disease or condition; improving a photopic (day light) vision; for improving correcting visual acuity, improving macular function, improving a visual field, or improving scotopic (night) vision by administration of retinal progenitor cells.

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.

A two-component hydrogel matrix system is provided, which system is useful in a variety of cell growth uses, including without limitation three-dimensional culture systems. The components comprise modified hyaluronic acid (HA); and modified elastin-like proteins (ELP). Variables of HELP, including for example, matrix stiffness, matrix stress relaxation rate, and cell-adhesive-ligand concentration, can be independently and quantitatively specified.