38 results about "Nonadherent cell" patented technology

The present invention provides a microorganism culture array, including one or more microflow cell culture array units, the microflow cell culture array unit includes a main channel and a plurality of cell culture units connected with the main channel, each culture unit includes a diffusion channel and a cell culture room, the cell culture room is connected with the main channel. The microorganism culture array of the invention not only realizes isolation of single-cell, but also makes the cell locate in zero flow state when the culture liquid or other reagent is changed, bringing greatly convenience for the cell culture and observation especially the culture and observation of non-adherent cells, greatly fit for research work of high throughput screen based on the cell phenotype.

The present invention provides a method for subculturing primate embryonic stem cells, and a method for inducing differentiation of the same cell into a vascular endothelial cell and a blood cell. The present invention provides a method comprising culturing primate embryonic stem cells in a medium containing a protein component without using feeder cells and cytokines in a container coated with an extracellular matrix, detaching colonies of the resulting embryonic stem cells in the presence of a cytodetachment agent, and plating the colonies in the similar medium, and a method comprising culturing primate embryonic stem cells in a serum-containing or not containing medium in the presence of cytokine, adhesion-culturing the resulting embryoid body or embroyid body-analogous cellular aggregate in the presence of a cytokine to obtain specific precursor cells, and separating non-adherent cells and adherent cells from the specific precursor cells to obtain blood cells and vascular endothelial precursor cells.

We have found a counter-intuitive way to improve the commercial-scale production of recombinant biological products in adherent-cell bioreactors, which reduces the risk of cell culture contamination, increases total yield and reduces the delay between seeding and harvest, thus minimizing expression product degradation, by inter alia inoculating an adherent culture bioreactor with suspension-adapted producer cells

The present invention relates to an apparatus for analyzing cells in real-time which incorporates a wavelength-tunable light source / infrared ray (IR) sensor and can be used to observe and analyze the IR-related characteristics of adherent cells or non-adherent cells. The Apparatus for analyzing cells in real-time in accordance with the present invention can be used to quantify specific materials in a cell and measure the metabolism of a cell. In addition, the apparatus for analyzing cells in real-time in accordance with the present invention can be configured to have a visible light microscope coupled thereto, and in this configuration, it can be used to locate a cell of interest.

The invention discloses a method used for separation and culture of poultry endothelial progenitor cells (EPCs). The method comprises following steps: peripheral blood is collected and separated so as to obtain mononuclear cells; the mononuclear cells are cultured in a culture solution; and a mixture of adherent cells and non-adherent cells is used for culture. It is possible to obtain a large amount of EPCs by separation and culture of poultry peripheral blood using the method.

Adherent cells bearing characteristics that are detectable only in the adherent state can be sorted on the basis of these characteristics independently of their adherent state, by applying a transformable label to the entire population of cells, both those bearing the characteristics of interest and those not, in their adherent state and identifying the locations of the cells of interest on the adherent surface. The cells of interest, or all cells other than those of interest, are then selectively treated to transform the labels and achieve differentiation between the cells of interest and the remaining cells. All cells are then released from the adherent state and sorted in the same manner as non-adherent cells but on the basis of whether the labels are transformed or not transformed.

A method of enriching stem or progenitor cells that includes growing a heterogeneous cell sample comprising stem and / or progenitor cells on a first substrate that is hydrophobic and has an elastic modulus less than about 100 MPa; recovering the heterogeneous cell sample from the first substrate; growing the recovered heterogeneous cell sample on a second substrate that is hydrophilic and has an elastic modulus higher than the elastic modulus of the first substrate to produce a subpopulation of nonadherent cells and a subpopulation of adherent cells; and recovering the nonadherent cell subpopulation, which is enriched for stem and / or progenitor cells. The invention also relates to a method of determining the clonogenic potential of a cell, such as a cancer stem cell enriched using the enrichment method described herein.

The invention provides a device, method, and system for high throughput detection of nucleic acid expression in individual cells. Cells are encapsulated in aqueous microdroplets which are merged with a biocompatible matrix, allowing on-chip fluorescence in situ hybridization on both adherent and non-adherent cells. The invention also provides multiplexed detection of nucleic acids, proteins, and cellular activity. The device and methods can be used to assess cellular interactions and to test the effects of antitumor agents.

The invention relates to methods of obtaining mesenchymal stem cells and is to provide a method of high-effectively obtaining mesenchymal stem cells from compact bone of mice. The method of high-effectively obtaining mesenchymal stem cells of mice includes the steps of: separating mesenchymal stem cells of mice, performing primary culture, and performing amplification culture to the mesenchymal stem cells of mice. The method greatly reduces the time of cell culture and can greatly increase quantity of produced cells, and is less in pollution of non-MSCs adherent cells. The invention, for the first time, discloses a multi-time bone sheet adhesion culture method, which can increase the yield of cells by 3-5 times. Through systematic researching on biological characteristics, such as growth status, surface markers, multidirectional differentiative potential and the like, of the MSCs from the compact bone of mice, it is proved that after 10 times of amplification culture in vitro, the cells still have strong characters of stem cells and can satisfy demands in experiment and researching. The method can quickly and high-effectively produce the stem cells. The obtained stem cells has high proliferation and growth speed and high differentiative potential.

the invention discloses a separating method of ox marrow mesenchyma stem cell, which comprises the following steps: adding ox marrow single-cell suspension liquid in the Percoll cell separating liquid with density at 1.082 to separate; obtaining foggy single-core cell at liquid intersecting layer; inoculating in the plastic container; removing non-adhesive wall cell to obtain the product with separating purifying efficiency over 98%.

Disclosed is a device for cell culture, the device includes a first chamber having an area for containing cells, such as isolated cells, for growth; a second chamber for containing culture medium; a semi permeable membrane forming at least part of a divider between the first chamber and the second chamber; wherein the area for containing cells for growth is adjustable via a moveable wall disposed within the first chamber. The device is especially suited for culturing of semi-adherent and non-adherent cells.

The invention discloses use of mesenchymal stem cells of dental pulp in obtaining of an artificial spinal cord and a preparation method for the artificial spinal cord. The preparation method consists of the following steps: (1) acquiring healthy deciduous teeth removed due to deformity correcting, digesting the healthy deciduous teeth with type 1 collagenase, inoculating a culture flask with the digested substance, carrying out washing to discard nonadherent cells, re-suspending adherent cells, then, carrying out culture, so as to obtain the mesenchymal stem cells of the dental pulp; (2) culturing the mesenchymal stem cells of the dental pulp with an alpha-MEM culture medium, so as to propagate the mesenchymal stem cells of the dental pulp; and (3) selecting third-generation dental-pulp mesenchymal stem cells, culturing the dental-pulp mesenchymal stem cells with an alpha-MEM culture medium containing vitamin C when the cell fusion rate is 90%so as to obtain a white membrane-shaped structure, and carrying out in-vitro molding, thereby finally obtaining the artificial spinal cord with a three-dimensional structure. The artificial spinal cord obtained by the preparation method can be used for inhibiting cicatrization and inflammation and promoting axon regeneration; and among rats suffering from complete spinal injury, Spinor therapy causes remarkable movement improvement, perception recovery and faster urination reflection recovery.

Nanostraws and to methods of utilizing them in order to deliver biologically relevant molecules such as DNA, RNA, proteins etc., into non-adherent cells such as immune cells, embryos, plant cells, bacteria, yeast etc. The methods described herein are repeatedly capable of delivering biologically relevant cargo into non-adherent cells, with high cell viability, dosage control, unaffected proliferation or cellular development, and with high efficiency. Among other uses, these new delivery methods will allow to scale pre-clinical cell reprogramming techniques to clinical applications.

The process of separating mesenchymal stem / ancestral cell from bone parenchyma includes the following steps: 1) digesting bone sclerite with collagenase; and 2) culturing the digested bone sclerite in culture medium, eliminating non-adherent cell and obtaining the adherent cell as the mesenchymal stem / ancestral cell. The process of the present invention is simple and practical, and the obtained mesenchymal stem / ancestral cell has high purity, no residual blood cell and endothelial cell and good differentiating capacity, and can differentiate to osteoplast, chondroblast, lipoblast, etc. The present invention establishes stable and practical mouse MSPC cell purifying culture technological system and lays foundation for the MSPC cell research.

The present invention relates to an apparatus for analyzing cells in real-time which incorporates a wavelength-tunable light source / infrared ray (IR) sensor and can be used to observe and analyze the IR-related characteristics of adherent cells or non-adherent cells. The Apparatus for analyzing cells in real-time in accordance with the present invention can be used to quantify specific materials in a cell and measure the metabolism of a cell. In addition, the apparatus for analyzing cells in real-time in accordance with the present invention can be configured to have a visible light microscope coupled thereto, and in this configuration, it can be used to locate a cell of interest.

The present invention provides a cell culture container for long term non-adherent cell culture which comprises having a coating containing a copolymer which is a copolymer having a recurring unit which contains a group represented by the following formula (a), a recurring unit which contains a group represented by the following formula (b), and a recurring unit which contains a group represented by the following formula (c), and a molar ratio of the recurring unit which contains a group represented by the formula (c) being 1 to 50 mol % based on the whole copolymer, provided with at least a part of a surface thereof[wherein, Ua1, Ua2, Ub1, Ub2 and Ub3, Rc, and An− are as defined in the specification and Claims.]

The invention provides a solution to the problem of transfecting non-adherent cells. Devices and delivery compositions containing ethanol and an isotonic salt solution are used for delivery of compounds and compositions to non-adherent cells.

The invention discloses a chip for evaluating mobile biological living body pharmacokinetics and a real-time detection system. The chip comprises a chip main body, at least one fluid flow passage arranged in the chip main body, a fluid outlet opened in the chip main body, and a passageway main body connecting a fluid inlet with the fluid outlet, wherein each fluid flow passage is provided with thefluid inlet which is opened in the chip main body; the passageway main body is internally provided with at least one referential chamber and at least one sample chamber, the sample chamber is provided with a liquid permeating protective screen which is vertically arranged in the passageway main body, and is arranged in the passageway main body through the liquid permeating protective screen in afluid communication way, the referential chamber and the sample chamber are separated from each other by a certain distance in space, and the referential chamber has the same shape and size as those of the sample chamber. The chip disclosed by the invention can be used for pharmacokinetics detection on fixed samples and non-fixed samples such as non-adherent cells, microorganisms and tiny organisms.

It is not understood what causes or influences pattern formation in cells during the development of an organism. When animal / human cells are cultured in a Petri dish the adherent cells attach to the bottom of the dish, whereas the non-adherent cells float in the growing medium. Currently there are no specialized dishes for culturing non-adherent cells. We now show that non-adherent cells could be induced to form distinct patterns when cultured in an etched plastic dish (Biosimulator). The non-adherent cells showed polarity when cultured in the etched plate. The polarity / pattern formation could be reversed with inhibitors specific for adhesion proteins. The phenomenon of pattern formation by non-adherent cells has wide applications in cell and developmental biology, diagnostics, microbiome research, biofluidics, drug discovery, industrial production of biological products, and also in biotechnology and bioengineering.

The invention relates to methods of obtaining mesenchymal stem cells and is to provide a method of high-effectively obtaining mesenchymal stem cells from compact bone of mice. The method of high-effectively obtaining mesenchymal stem cells of mice includes the steps of: separating mesenchymal stem cells of mice, performing primary culture, and performing amplification culture to the mesenchymal stem cells of mice. The method greatly reduces the time of cell culture and can greatly increase quantity of produced cells, and is less in pollution of non-MSCs adherent cells. The invention, for the first time, discloses a multi-time bone sheet adhesion culture method, which can increase the yield of cells by 3-5 times. Through systematic researching on biological characteristics, such as growth status, surface markers, multidirectional differentiative potential and the like, of the MSCs from the compact bone of mice, it is proved that after 10 times of amplification culture in vitro, the cells still have strong characters of stem cells and can satisfy demands in experiment and researching. The method can quickly and high-effectively produce the stem cells. The obtained stem cells has high proliferation and growth speed and high differentiative potential.

Nanostraws and to methods of utilizing them in order to deliver biologically relevant molecules such as DNA, RNA, proteins etc., into non-adherent cells such as immune cells, embryos, plant cells, bacteria, yeast etc. The methods described herein are repeatedly capable of delivering biologically relevant cargo into non-adherent cells, with high cell viability, dosage control, unaffected proliferation or cellular development, and with high efficiency. Among other uses, these new delivery methods will allow to scale pre-clinical cell reprogramming techniques to clinical applications.

The present invention relates to a method for separating and identifying human placental myofibroblasts. After the placental fetal decidual tissue was mechanically disrupted, it was digested with DMEM digested solution containing collagenase I, and isolated into mononuclear cells. Cultivate under humidity to induce adherence. After 48 hours, change the medium in full to remove non-adherent cells and add fresh medium. After adherent cells form monoclonal cells, each monoclonal cell is cultured separately. When the cells are about 90% confluent, Trypsin passage to obtain myofibroblasts. The invention adopts the combined method of cell morphology, cell cycle, cell surface marker detection, cell adipogenic and osteogenic differentiation ability detection, cell pluripotent stem cell factor detection and cell desmin expression detection to identify the obtained myofibroblasts.

Conventional dishes and plates do not induce adhesion of non-adherent eukaryotic cells and microorganisms. When animal / human cells are cultured in a Petri dish the adherent cells attach to the bottom of the dish, whereas the non-adherent cells float in the growing medium. Less than 50% of the microorganisms are culturable using conventional techniques. Currently there are no specialized dishes for culturing non-adherent cells or microorganisms. A method to induce adhesion of non-adherent eukaryotic and prokaryotic cells involves use of engravings on plastic or metal surfaces (Biosimulator). The engravings also induce proliferation of microorganisms. The non-adherent cells and microorganisms show polarity when cultured in the engraved plate. The polarity / pattern formation could be reversed with inhibitors specific for adhesion proteins. Induction of adherence and proliferation on an engraved surface has wide applications in cell and developmental biology, diagnostics, microbiome identification, biofluidics, drug discovery, industrial production of biological products, and also in biotechnology and bioengineering.

The invention provides a solution to the problem of transfecting non-adherent cells. Methods and compositions containing ethanol and an isotonic salt solution are used for delivery of compounds and compositions to non-adherent cells, e.g. T cells.

The invention discloses a method for isolating and culturing poultry endothelial progenitor cells, which is characterized by comprising the following steps: collecting peripheral blood, separating and collecting mononuclear cells, adding culture medium for cell culture, using adherent cells and non-adherent cells The mixture is cultivated. According to the method for isolating and culturing poultry endothelial progenitor cells of the present invention, a large amount of EPCs can be isolated and cultured from poultry peripheral blood.

A method of enriching stem or progenitor cells that includes growing a heterogeneous cell sample comprising stem and / or progenitor cells on a first substrate that is hydrophobic and has an elastic modulus less than about 100 MPa; recovering the heterogeneous cell sample from the first substrate; growing the recovered heterogeneous cell sample on a second substrate that is hydrophilic and has an elastic modulus higher than the elastic modulus of the first substrate to produce a subpopulation of nonadherent cells and a subpopulation of adherent cells; and recovering the nonadherent cell subpopulation, which is enriched for stem and / or progenitor cells.

The invention relates to a separation and identification method of human placenta myofibroblasts. The separation and identification method comprises the following steps of: mechanically crushing the decidual tissue of the placenta fetal surface, digesting the decidual tissue by DMEM digestive juice containing collagenase I and thereby separating the decidual tissue into individual karyocytes; cultivating the karyocytes by using a DMEM / F12 culture medium containing 10% of fetal calf serum under the conditions of 37 DEG C, 5% of CO2 and saturated humidity; inducing adherence to wall and completely replacing the liquid in 48 hours, removing cells not adhering to the wall, adding fresh culture medium, and after the cells adhering to the wall is mono-cloned, cultivating each monoclonal cell separately; and when the cells are fused by about 90%, performing pancreatin passage, thereby obtaining the myofibroblasts. The method further identifies the obtained myofibroblasts by the combined method of cell morphology, cell cycle, cell surface marker detection, cell adipogenesis and osteogenesis differentiative capacity detection, cell multipotential stem cell factor detection and cell desmin expression detection.

The invention provides a device, method, and system for high throughput detection of nucleic acid expression in individual cells. Cells are encapsulated in aqueous microdroplets which are merged with a biocompatible matrix, allowing on-chip fluorescence in situ hybridization on both adherent and non-adherent cells. The invention also provides multiplexed detection of nucleic acids, proteins, and cellular activity. The device and methods can be used to assess cellular interactions and to test the effects of antitumor agents.

The invention relates to a preparation method of mesenchymal stem cells. The preparation method comprises the following steps of (1) collecting the same parts of bone marrow (BM) aspiration substancesfrom thigh bones and shin bones of a green fluorescent protein transgenic mouse under the aseptic condition; (2) selecting bone marrow mononuclear cells by a Ficoll-Paque density gradient centrifugation; (3) performing culturing in improved EAG culture mediums (MEM-alpha) to which 20vol% of fetal calf serum (FBS), 100[mu]l / ml of penicillin and 100[mu]g / ml of streptomycin are supplemented, in a flask of 75-cm<2> under the condition of 37 DEG C and 5% of CO2; (4) replacing culture mediums containing non-adherent cells once every 4-5 days; and (5) when 90% confluence is achieved, harvesting adherent cells through trypsin digestion, wherein under the condition, Sca-1 positive cells exceed 96.67%. According to the preparation method disclosed by the invention, the cell proliferation speed canbe substantially increased, ample nutrition is provided for cells, protein synthesis is promoted, and the treatment effects of treating systemic lupus erythematosus through stem cells can be effectively increased and reinforced.