A kit, including: a first solution, a second solution, and a metal mesh. The first solution includes: dulbecco's modified eagle medium (DMEM), 65-95 V/V %; dimethyl sulfoxide (DMSO), 5.5-20 V/V %; ethylene glycol (EG), 3.5-15 V/V %; bovine serum albumin (BSA), 0.5-4 W/V %; sucrose 1-5 W/V %; methylcellulose with a viscosity of 4000 centipoise (cP), 0.05-0.8 W/V %; hetastarch 0.25-0.6 W/V %; and glucose 15-35 W/V %. The second solution includes: DMEM, 65-95 V/V %; DMSO, 5.5-20 V/V %; EG, 8-20 V/V %; BSA, 0.5-4 W/V %; sucrose, 10-20 W/V %; methylcellulose with a viscosity of 4000 centipoise (cP), 0.05-0.8 W/V %; polyvinyl pyrrolidone (PVP), 0.25-0.6 W/V %; and glucose 15-35 W/V %. The metal mesh has a thickness of 0.15-0.2 mm and includes a plurality of square holes. The side length of the square holes is 2.0-3.0 mm, and the spacing between adjacent holes is 0.5-2.0 mm.

Provided herein are composite scaffold biomaterials including two or more scaffold biomaterial subunits, each including a decellularized plant or fungal tissue from which cellular materials and nucleic acids of the tissue are removed, the decellularized plant or fungal tissue having a 3-dimensional porous structure, the two or more scaffold biomaterial subunits being assembled into the composite scaffold biomaterial and held together via gel casting using a hydrogel glue; via complementary interlocking geometry of the two or more scaffold biomaterial subunits; via guided assembly based biolithography (GAB); via chemical cross-linking; or any combinations thereof. Methods for producing such scaffold biomaterials, as well as methods and uses thereof, are also provided.

A method of circulating tumor cells isolation, using an isolating cultural system of circulating tumor cells, comprises the following steps: (1) providing a sample; (2) adding a cell culture medium to the isolating cultural system of circulating tumor cells; (3) adding the sample to the isolating cultural system of circulating tumor cells to cultivate; and (4) collecting the suspended circulating tumor cells in the cell culture medium; wherein the isolating cultural system of circulating tumor cells comprises a container including a cell adhesion portion, and cellulose coated on the cell adhesion portion.

The present disclosure provides a method for freeze-drying cells in a hydrogel comprising nanofibrillar cellulose, the method comprising providing a hydrogel comprising nanofibrillar cellulose, providing cells, combining the cells and the hydrogel comprising nanofibrillar cellulose to form a cell system, and freeze drying the cell system to obtain dried cells in a hydrogel comprising nanofibrillar cellulose. The present disclosure also provides a freeze-dried hydrogel comprising nanofibrillar cellulose and cells.

The present application provides a method for preparing transplantable cell preparation, the method comprising culturing eukaryotic cells at conditions allowing the cells to coalesce and form cell aggregates, providing the cell aggregates in a nanofibrillar cellulose hydrogel to obtain a transplantable cell composition comprising eukaryotic cells in a nanofibrillar cellulose hydrogel matrix, and a transplantable cell preparation. The present application also provides the transplantable cell composition for use in a therapeutic method, and to use of nanofibrillar cellulose for preparing the transplantable cell composition.

The present disclosure relates to agarose and methylcellulose storage and transport mediums, systems for cell storage and transport, and cell storage and transport methods. The instantly-disclosed agarose and methylcellulose storage and transport medium is ideally suited for 3D spheroid cell culture storage and transport. In particular aspects, the storage and transport mediums, systems, and methods are used in combination with or performed in labware that combine 3D spheroid culture with a gas permeable, micro-patterned design that allows for protection and prolonged maintenance of spheroid cell (e.g. hepatocytes) viability and functionality during storage and transport.

Standardized nanocellulose 3D matrices for in vitro human and animal cell culture with batch-to-batch regularity in terms of porosity on both surfaces, measured in percent, as well as elasticity, measured by Young's Modulus. A method of manufacturing these 3D nanocellulose matrices, in addition to the matrices manufactured by this method, modified for obtaining nanocellulose 3D matrices containing distinct distribution of nanofibers on the matrix surfaces, considering or not the immobilization, absorption or adsorption of other chemical molecules, resulting in bioengineered physical, chemical, biological and mechanical properties for obtaining an in vitro platform to be used in the cultivation of human and animal cells where the behavior of these cells is evaluated on a time scale (4D). The present invention further encompasses the use of these bioengineered nanocellulose 3D matrices in the development of reconstructed artificial skin in the laboratory with the intention of serving as a platform for testing the efficacy and safety of cosmetics and drugs in vitro, as a platform for in vitro culture of animal and human cells, as a 3D platform for in vitro cytotoxicity and genotoxicity testing, as a platform for in vitro fertilization.

Methods and compositions for promoting donor-specific tolerance and immunocompetence to a recipient of a solid organ transplant, by implanting an allogeneic solid organ in a recipient in need of a solid organ transplant and further comprising surgical implantation of a tissue-engineered allogeneic cultured postnatal thymus tissue product in the recipient of a solid organ from a donor.

Methods and systems for assessing membrane potential are provided. In some embodiments, the methods and systems, described herein, may allow spatial patterns of membrane potential to be facilely obtained. For instance, a method may comprise transferring a population of cells from a tissue to a substrate. The transfer process may substantially maintain the viability of and/or the spatial relationship between the cells. The cells on the membrane may be exposed to a voltage sensitive dye. The dye may allow the membrane potential of individual cells on the substrate to be imaged or otherwise detected. The individual cell membrane potentials when imaged together on the substrate may form a spatial membrane potential pattern. The spatial membrane potential pattern may be used to assess one or more physiological characteristics of the cells. The methods and systems may be used for a wide variety of applications, including the assessment of biopsies.

The invention discloses a method of manufacturing polysaccharide beads, comprising the steps of: i) providing a water phase comprising an aqueous solution of a polysaccharide; ii) providing an oil phase comprising at least one water-immiscible organic solvent and at least one oil-soluble emulsifier; iii) emulsifying the water phase in the oil phase to form a water-in-oil (w/o) emulsion; and iv) inducing solidification of the water phase in the w/o emulsion, wherein the organic solvent is an aliphatic or alicyclic ketone or ether.