Provided herein are methods of perfusion culturing an adherent mammalian cell using a shake flask and a plurality of microcarriers, and various methods that utilize these culturing methods.

A microcarrier for cell culture and expansion is provided. The microcarrier includes decellularized mammalian tissue. Further, the microcarrier has an average particle size ranging from about 10 micrometers to about 600 micrometers. A method of forming a decellularized mammalian tissue microcarrier for cell culture and expansion is also provided, along with a method for treating a mammalian tissue defect via a decellularized mammalian tissue microcarrier on which cells from the same tissue type as the decellularized mammalian tissue are expanded.

The present invention is directed to a method of producing compositions including embryonic proteins. The method includes culturing cells under hypoxic conditions on a biocompatible surface in vitro. The culturing method produces both soluble and non-soluble fractions, which may be used separately or in combination to obtain physiologically acceptable compositions useful in a variety of medical and therapeutic applications.

Aspects of the present disclosure include methods that include co-culturing a cell and a microparticle that includes a capture ligand, in a culture medium under conditions in which a biomarker produced by the cell is bound by the capture ligand. Such methods may further include detecting (e.g., by flow or mass cytometry) complexes that include the microparticle, the capture ligand, the biomarker, and a detection reagent. The methods may further include determining the proportion or number of cells among a heterogeneous cell population that produced the biomarker and/or the level of biomarker secreted by such cells. Compositions, systems and kits are also provided.

Methods of repeated aggregate dissociation and reformation of pluripotent stem cells (PSCs) within the same bioreactor until a desired final cell number is achieved. A preferred step-wise process for controlled growth of PSCs and aggregate size using periodic dissociation with a dissociation medium which contains either proteolytic enzymes or chemical reagents, mechanical agitation, or a combination of these methods.

The present invention provides a method polyclonal stimulation of T cells, the method comprising contacting a population of T cells with a nanomatrix, the nanomatrix comprising a) a flexible matrix, wherein said matrix is of polymeric material; and b) attached to said polymeric flexible matrix one or more polyclonal stimulatory agents which provide activation signals to the T cells; thereby activating and inducing the T cells to proliferate; wherein the nanomatrix is 1 to 500 nm in size. At least one first and one second stimulatory agents are attached to the same or to separate flexible matrices. If the stimulatory agents are attached to separate beads, fine-tuning of nanomatrices for the stimulation of the T cells is possible.

The invention relates to a matrix in ball form comprising cross-linked fibrinogen, the matrix being free from fibrin, as well as to a method for preparing such a matrix, comprising the following steps: (a) providing an initial composition comprising fibrinogen and a platelet factor; (b) injecting said initial composition into an oil heated to a temperature of 50 C. to 80 C. so as to form an emulsion; (c) mixing the emulsion thus obtained at a temperature of 50 C. to 80 C. until a matrix in ball form is obtained; and (d) isolating the matrix thus obtained. The matrix is used as a cell carrier.

A closed, automated and scalable stirred tank bioreactor platform, capable of sustaining high fold expansion of hPSCs is provided. hPSCs are expanded in a controlled bioreactor using perfused xeno-free media. Cell harvest and concentration are performed in closed steps. The hPSCs can be cryopreserved to generate a bank of cells or further processed as needed. Cryopreserved cells can be thawed into a 2D tissue culture platform or a 3D bioreactor to initiate a new expansion phase or be differentiated to the clinically relevant cell type. The expanded hPSCs express hPSC-specific markers, have a normal karyotype and the ability to differentiate to the cells of the three germ layers. This end-to-end platform allows large expansion of high quality hPSCs that can support the required cell demand for various clinical indications.

An in vitro analysis diagnostic instrument, and a microfluidic chip and method for sorting and enriching circulating tumor cells. The microfluidic chip includes a functional board. A first side face of the functional board is provided with a primary sorting channel in communication with a sample inlet and a fine screening channel in communication with the primary sorting channel, and a deepened channel is dug on the side of the fine screening channel away from the aggregation of circulating tumor cells. The deepened channel is arranged in an extension direction of the fine screening channel, and the deepened channel has a depth greater than that of the fine screening channel.

There is provided a method for producing hydrogel microparticles with spherical shape and having a narrow-disperse or mono-disperse size distribution. At least one temporary crosslinker such as those of formula (I), (Ila)-(Ilf) and at least one permanent crosslinker comprising two or more vinyl groups, such as: divinylbenzene (DVB), ethylene glycol dimethacrylate (EGDMA),diethyleneglycol dimethacrylate (DEGDMA), N,N-methylenebisacrylamide (MBA), oligo/poly ethyleneglycol dimethyacrylate, 1,4-butanediol dimethacrylate, and 1,6-hexanediol dimethacrylate are combined in an organic solvent having a polarity suitable for a precipitation polymerization to occur. The precipitation polymerization is allowed to take place without the addition of surfactant and/or stabilizer and/or the formed microparticles comprise less than 1% surfactant and/or stabilizer. These microparticles may be further functionalized to obtain amine and carboxylic acid units by functionalizing the monomers of the temporary crosslinkers. The functionalized microparticles are used for cryopreserving cells or as a vaccine delivery platform.

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