192 results about "Cell bank" patented technology

The invention discloses a method for constructing a human peripheral blood immune cell bank. The method comprises the following steps of: collecting human peripheral blood, separating autologous plasma, separating a peripheral blood mononuclear cell, separating a mononuclear cell by the peripheral blood mononuclear cell and freezing, separating a T lymphocyte by the peripheral blood mononuclear cell and freezing, separating a B lymphocyte by the peripheral blood mononuclear cell and freezing, separating an NK cell by the peripheral blood mononuclear cell and freezing, and encoding and puttingin storage. According to the invention, immune cells of health or young people are separated and are respectively independently frozen, and relative numbers are stored and put into storage, so that the stored human immune cells have the characteristics of high activity, high purity and convenience for use, and fetal calf serum can be replaced by the human autologous plasma, so that introduction of a foreign protein can be avoided. Meanwhile, the method, provided by the invention, has the advantages of low cost, low requirements on laboratory conditions, and wide application.

A mixed-height cell library for designing integrated circuits is provided. The mixed-height cell library includes a first plurality of cells having a first track height and a second plurality of cells having a second track height that are configured to be coupled to the first plurality of cells at respective power and ground rail lines. A method for mixed-height cell placement and optimization is also provided. The method comprises abutting cells of different track heights to form a plurality of rows of cells by coupling power and ground rails of the cells at a secondary layer that is different from a primary layer that is used to connect active material and determining whether re-ordering cells within rows allows for further compaction of adjacent rows. The method further comprises re-ordering cells within rows so to allow for further compaction of adjacent rows. The method also includes the steps of splitting rows vertically to minimize the distance between the split rows.

The invention provides a humanized PD-L1 tumor cell line MC-38-hPD-L1, a builtanimal tumor model with the same and a method for constructing the humanized PD-L1 tumor cell line. The method particularly includes knocking out animal-origin PD-L1 by the aid of CRISPR-CAS9; carrying out amplification and cultivation to obtain knocked-out cell banks; extracting DNA (deoxyribonucleic acid) and carrying out PCR (polymerase chain reaction) amplification; recycling and cloning amplification products; carrying out over-expression on human-origin PD-L1 in MC-38 cell lines of mPD-L1 KO by the aid of lentivirus systems; packaging lentivirus and screening Puromycin to obtain the humanized MC-38 cell line of PD-L1. The humanized PD-L1 tumor cell line, the animal tumor model and the method have the advantages that as shown by results, high killing efficiency and multiplication capacity are obviously presented by tumor infiltration CD8 T lymphocytes after antibody treatment is carried out, tumor infiltration Treg cells can be obviously inhibited after antibody treatment is carried out, and accordingly the method is proved to be effective and feasible from the aspect of molecular mechanisms.

Methods and systems are provided for analyzing cells of a cell library used to generate a layout. One exemplary method involves determining a routed connection location utilized in the layout for a pin of the cell for each instance of the cell in the layout. The method continues by determining a utilization metric for the pin of the cell based on the plurality of routed connection locations and a plurality of possible connection locations for the pin, and displaying the utilization metric on a display device.

The present invention discloses a method of isolation, pooling and further culturing of Mesenchymal Stem cells (MSC) for clinical application. Present invention also discloses the method of establishing Master Cell bank, followed by Working Cell Bank from which the final therapeutic composition referred to as Investigational Product / Investigational Medicinal Product comprising of allogenic bone marrow-derived MSC is formulated for clinical applications.

A method for constructing banks of human dental pulp stem cells comprises the steps of acquiring basic information, obtaining dental pulp stem cells, cryopreserving and putting dental pulp stem cells into cell banks and recording and checking information. When the banks of dental pulp stem cells are used, recovery and multiplication culture are carried out in accordance with the actual demands. In the method, the dental pulp stem cells are obtained from dental pulp of waste healthy human teeth and a reserve bank for effective resources is established by using system engineering. A great quantity of dental pulp stem cells with functional activities can be obtained through short-term culture of the human dental pulp stem cells stored in the banks and can be stored for long term without losing the activities. Compared with the prior art, the invention is characterized in that the construction operation specification is simple, easy to grasp, safe and feasible; the bank construction cost is low and the standardization degree is high; and the cells from different age groups adopt different cryopreservation solution so that the activities of the recovered cells are enhanced, thus expanding the sources of the cells and enhancing the activities of the cells. The teeth used in the method are those which are extracted for various reasons and are waste, thus greatly saving the medical resources; therefore, the construction has wide application prospect.

The invention relates to the technical field of cell preservation, in particular to a storage method of immune cells and a cell freezing medium. The method includes: separating and collecting peripheral blood mononuclear cells, and adding the cell freezing medium to store the cells in a frozen manner, wherein the cell freezing medium comprises, by weight percentage, 0-62% of serum-free liquid culture medium, 0.5-1.5% of penicillin-streptomycin solution, 5-10% of dimethyl sulfoxide and 30-92% of autologous plasma. The storage method of the immune cells has the advantages that the method is simple and effective, the infectious disease risks of exogenous serum are avoided, the survival rate of the revived freezing-stored cells after induction culture is above 90%, the various biological indexes of the cells satisfy the requirements, and a good storage effect is achieved; the method can monitor and trace cell bank samples in real time, accurately record information such as cell storage positions, temperature and cell sources, each cell has a unique identifier, and the method is convenient to use and safe and reliable.

The present invention discloses a method of isolation, pooling and further culturing of Mesenchymal Stem cells (MSC) for clinical application. Present invention also discloses the method of establishing Master Cell bank, followed by Working Cell Bank from which the final therapeutic composition referred to as Investigational Product / Investigational Medicinal Product comprising of allogenic bone marrow-derived MSC is formulated for clinical applications. Present disclosure also discloses a robust manufacturing process for consistent production of clinical grade Mesenchymal Stromal cells (MSCs). The process enables production of highly viable potent cells. The process steps relating to preparation of media, cell seeding, harvesting are fine tuned to achieve consistency in cell yield, superior cell viability, purity, improved cell proliferation, high cell recovery, low HLA-DR expression, reduction in culture duration. The viability and purity of cells are further improved by optimized wash process without cell loss / cell stress. The disclosure further provides a method of cyrostoring MSCs at high cell density without affecting the viability of cells. It further provides economical means to store and transport at −80° C.

The invention discloses a preparation method of human adipose-derived MSCs (mesenchymal stem cells) and an application of the human adipose-derived MSCs in preparation of medicine for treating diseases. The preparation method of the human adipose-derived MSCs comprises steps as follows: adipose tissue collection, adipose tissue transportation, adipose tissue connection, adipose tissue microblock preparation, SVF (stromal vascular fraction) extraction, primary culture, subculturing, cell cryopreservation, cell bank establishment and cell recovery. The human adipose-derived MSCs have a powerful immunity regulation function and can be applied to preparation of medicine for treating autoimmune diseases, and compared with conventional medicine, the human adipose-derived MSCs have the advantages as follows: 1, the treatment is convenient, the curative effect is lasting, the variety and the number of medicine for treatment and toxic and side effects caused by medicine treatment can be reduced, even the medicine such as immunosuppressant and the like for treatment is out of service, the death rate and the disability rate are reduced, and life quality is improved; 2, the safety is good, and toxic and side effects are avoided; 3, the source of adipose tissue is wide, the collection is convenient, fundamental research of the human adipose-derived MSCs is solid, the preparation technology is mature, the popularization degree is high, and the technology is controllable; and 4, the preparation cost of the human adipose-derived MSCs is lower than that of other biological preparation and can be accepted by most patients.

The invention provides methods for treating pathological conditions associated with an undesirable inflammatory component. The invention is generally directed to reducing inflammation by administering cells that have one or more of the following effects in an injured subject: interact with splenocytes, preserve splenic mass, increase proliferation of CD4+ and CD8+ T-cells, increase IL-4 and IL-10, decrease IL-6 and IL-1β, and increase M2:M1 macrophage ratio at the site of injury. The invention is also directed to drug discovery methods to screen for agents that modulate the ability of the cells to have these effects. The invention is also directed to cell banks that can be used to provide cells for administration to a subject, the banks comprising cells having desired potency for achieving these effects.

The invention discloses a culture method for largely amplifying hair follicle stem cells in vitro, which belongs to the technical field of cell culture. The invention establishes a method for largely amplifying hair follicle stem cells in vitro, which uses microspheres as carriers for cell culture and a rotary bottle for cell proliferation fermentation tank. The method is simple and convenient in operation, economic and practical, avoids reduction in cell proliferation capacity of by repeated passage cells, differentiation potential and the like and related side effects of the conventional cell amplification method and saves culture solution at the same time; the hair follicle stem cells cultured by the method have high multiplication capacity and retain original differentiation potential and can be used for: (1) building hair follicle stem cell bank for providing high-quality seed cells for study on adult stem cells; (2) transplanting and repairing pathological tissue organs; (3) constructing tissue-engineered organs which are used as substitutes for autologous organs for transplanting and repairing pathological or loss tissue organs; and (4) serving as target cells in genetic treatment for treating corresponding diseases.

The invention relates to a construction method of an immune cell bank. The construction method comprises the following steps: processing placenta blood or placenta tissue obtained from umbilical cord blood, peripheral blood and placenta tissue so as to obtain mononuclear cells; conducting amplification culture on the mononuclear cells, or conducting amplification culture on the mononuclear cells after freezing the mononuclear cells; and coding the obtained immune cells and storing the immune cells. The construction method disclosed by the invention, on the basis of achieving the efficient amplification of the immune cells, can still keep the frozen immune cells at high activity and high purity; moreover, the resuscitation activity of the stored immune cells achieves above 85%, the immune cells are resurrected within 30-60min and the storage duration is 15-20 years; the application scope of blood storage and tissue storage to the treatment by virtue of three categories of medical technologies is enhanced, and a novel and healthy storage mode is provided for the prevention and the treatment of tumors and for the prevention and the treatment of immune system diseases caused by virus infection.

The invention provides a method for establishing a human umbilical cord mesenchymal stem cell bank by adopting a serum substituent and relates to a method for establishing a stem cell bank. The method comprises the following steps: separating umbilical cord Wharton jelly and adding into a serum-free culture medium for culturing; replacing the culture medium for one time every 5 days; after cells grow to 70 percent, reproducing; collecting tissue blocks again and adding into the serum-free culture medium for secondary culture; replacing the culture medium for one time every 5 days; after cells grow to 70 percent, reproducing; after subculture, cryopreserving to establish the umbilical cord mesenchymal stem cell bank. The invention adopts serum-free culture and a cryopreserving system to establish the cell bank and a tissue block secondary climbing method. The method has the advantages that all tissues are effectively utilized and the cells can climb out as much as possible; the quantity of obtained cells is twice of that of a traditional method. A cell culture system adopts the serum substituent to replace bovine serum and adopts a pancreatin substituent to replace porcine pancreatin, so that pollution to human by animal sources is effectively avoided, and a quality control link of detecting cattle and pigs is saved.

The invention discloses a method for obtaining human adipose-derived stem cells and a construction method for a multilevel allogeneic adipose-derived stem cell bank, relates to an adipose-derived stem cell obtaining method and an adipose-derived stem cell bank construction method, and aims to solve the problems of great damage to cell activity caused by a fat collection method and a conventional human adipose-derived stem cell obtaining method and high aging rate of stem cells used by multiple persons after subculture of many times in a conventional adipose-derived stem cell bank. The method for obtaining the stem cells comprises the following steps of 1, screening donors; 2, collecting fats; 3, transporting the fats; 4, separating the human adipose-derived stem cells. The construction method for the stem cell bank comprises the following steps of 1, screening the donors; 2, collecting the fats; 3, transporting the fats; 4, separating the human adipose-derived stem cells; 5, culturing, subculturing and amplifying the human adipose-derived stem cells; 6, constructing a main cell bank; 7, establishing working cell banks. According to the methods, the cell activity is less damaged, and multiple levels of stem cell banks for multiple persons are established for the establishment of the adipose-derived stem cell bank.

The invention discloses a serum-free cell cryopreservation solution. The serum-free cell cryopreservation solution is prepared from amino acid, inorganic salt, albumin, extracellular cryoprotectant and the like. The serum-free cell cryopreservation solution contains double protective components and can synchronously protect cells from being damaged by ice crystals from cell interiors to cell exteriors during cryopreservation, the recovery rate of the cells is greatly improved, and the serum-free cell cryopreservation solution is suitable for preservation of wide cell lines, wherein the serum-free cell cryopreservation solution can be used in long-term cell preservation, cell bank establishment, scientific-research cell strain preservation and long-term preservation of clinical mesenchymal stem cells, mononuclear cells, immune cells and all related mammal cells; meanwhile, the serum-free cell cryopreservation solution is a cryopreservation solution completely without serum, the situation is avoided that heterogeneous and exogenous components are introduced in clinical application, and the serum-free cell cryopreservation solution is safer.

The invention provides a construction method of a human adipose derived stem cell bank. The construction method comprises the following steps: (1) collecting adipose tissues, adding a saline solution and centrifuging; (2) removing an oil layer and washing the adipose tissues; (3) inoculating the washed adipose tissues into a complete culture medium and culturing under the conditions that the temperature is 37 DEG C and the concentration of CO2 is 5 percent; (4) replacing the culture medium for one time every 5 to 8 days; when the cell confluence is 70 percent to 80 percent, digesting cells with TrypLETM Select; (5) carrying out subculture; (6) detecting the quality; (7) cryopreserving; (8) establishing the bank. According to the construction method provided by the invention, oil and red blood cells are removed by adopting a centrifugal method and collagenase is not used for digesting; the operation time can be shortened and a process is simple and feasible; a lot of the red blood cells are not generated, and the red blood cells do not need to be removed by adopting a red blood cell lysis solution or low-molecular-weight heparin calcium; potential risks on human adipose derived stem cells are not caused; meanwhile, the cost is saved.

An in vitro method is provided for analyzing chromatin accessibility and screening RNA of each single cell in a heterologous population (e.g., a library of cells). The method comprises incubating cell nuclei obtained from lysed cells with a transposome complex in a tagmentation buffer, performing reverse transcription wherein each of the RNAs is reverse transcribed to a DNA barcoded with the first barcode; sequencing DNA, which is extracted from digested cell nuclei; and analyzing chromatin accessibility and RNA of the cells. In a further embodiment, the method described comprises performing combinatorial cellular indexing and / or a perturbation step. Additionally, provided are a transposase TnY, buffer(s), and kit(s) for use in the described method.

Techniques for organizing a cell library permit a large number of cells. To improve design accuracy using cell libraries, very large cell libraries are needed. However, optimization tools are not able to use very large cell libraries directly, since their results suffer. Very large cell libraries are organized into sublibraries that are adapted to be processed by optimization tools. This allows improvement in the design quality of integrated circuits, while allowing the designs to be processed by optimization tools.

The invention provides a synovial sarcoma cell line hss-005R, which is preserved in the China Center for Type Culture Collection (CCTCC) with the preservation number of CCTCC NO. C201911. The invention further provides an offspring cell line of the synovial sarcoma cell line hss-005R. A new human synovial sarcoma cell line is established, which is stable in character and can be stably passed for multiple generations. Under the premise of retaining main clinical biological characteristics, the established human synovial sarcoma cell line has the characteristics of high tumor formation rate, short incubation period, good uniformity and the like, enriches a synovial sarcoma cell bank, and can successfully establish a synovial sarcoma animal model, wherein the established animal model can be used for fundamental research and drug screening, so as to provide powerful scientific research data for the research developed on the basis of Chinese population genetic background, and provide new test material for test on sensibility and tolerance of a clinical anti-cancer drug of a new drug preclinical study in vivo experiment.

The invention provides methods for treating traumatic brain injury. The invention is generally directed to treating traumatic brain injury by administering cells that have one or more of the following effects in an injured subject: interact with splenocytes, preserve splenic mass, increase proliferation of CD4+ and CD8+ T-cells, increase IL-4 and IL-10, and increase M2:M1 macrophage ratio at the site of injury. The invention is also directed to drug discovery methods to screen for agents that modulate the ability of the cells to have these effects. The invention is also directed to cell banks that can be used to provide cells for administration to a subject, the banks comprising cells having desired potency for achieving these effects.

Compositions and methods of treating mammalian diseases using myoblasts, and / or their physical, genetic, chemical derivatives. Myogenic cells that are normal, or genetically or phenotypically altered are cultured and transplanted into malfunctioning and / or degenerative tissues or organs to alleviate conditions that are hereditary, degenerative, debilitating, undesirable, and / or fatal. Treatment of these conditions is not limited to the usage of mechanical, electrical or physical properties of these myogenic cells, but includes the usage of biochemicals secreted / released by the latter. The present invention discloses the use of normal myoblasts to deliver the complete normal genome to effect genetic repair, or to augment the size, or the function of tissues or organs. Certain conditions may be better served with genetically altered myogenic cells derived from gene transduction, whereas others may be better served with cytoclimes converter cells. Endogenous biochemical(s) are used to control cell fusion of myoblasts among themselves or with other cell types. An automated cell processor within a cell bank which enables the manufacture, at a single run, of unprecedented large quantities (greater than 100 billion) of normal or genotypically or phenotypically altered myogenic cells is also disclosed.

The invention discloses a serum-free cell cryopreservation solution which comprises an amino acid component, an inorganic salt component, an extracellular cryoprotectant, an intracellular cryoprotectant and the like. The cell cryopreservation solution disclosed by the invention has the advantages that double protective components are contained and a cell is simultaneously protected from ice crystal damage during cryopreservation intracellularly and extracellularly, so that the recovery rate of the cell is greatly increased; the cell cryopreservation solution is suitable for extensive cell linepreservation, can be used for long-term preservation of cells and can be applied to establishment of cell banks, scientific cell line preservation and long-term preservation of clinical mesenchymal stem cells, monocytes, immune cells and all mammalian cells involved; and meanwhile, the cell cryopreservation solution is a completely serum-free, protein-free and dimethylsulfoxide (DMSO)-free cryopreservation solution, so that heterologous exogenous components introduced in clinical applications are avoided and the solution is safer.

The invention relates to an efficient-secretion canine parvovirus (CPV) resistant monoclonal antibody hybridoma cell A135 strain and belongs to the technical field of biology. The hybridoma cell A135 strain selected from the secretion CPV resistant monoclonal antibody hybridoma cell bank is excellent in biological performance and is injected with ascetic fluid generated in a BALB / C mouse peritoneal, so that neutralizing titer is up to 1010. In addition, the hybridoma cell A135 strain has moderate neutralizing capacity for various CPV subtype virus strain such as CPV-2a, CPV-2b, CPV-2c (a) and CPV-2c (b) and CPV strains from foxes and raccoon dogs, is wide in anti-CPV strain range and is used for clinical treatment of attacked dogs with CPV diseases, and effective rate is up to 100%.

An automatic layout apparatus is provided with: a storage device storing a cell library containing therein cell library data; and a layout tool obtaining from the cell library the cell library data associated with cells to be placed as described in a netlist to perform automatic placement of the cells to be placed. The obtained cell library data include layout coordinates of diffusion layers within the cells to be placed. The layout tool determines positions of the cells to be placed, referring to the layout coordinates of the diffusion layers.

A human umbilical cord blood immune cell bank establishing method comprises the steps that umbilical cord blood collection and transportation are performed; a mononuclear cell is separated from umbilical cord blood; a CD34+ cell in the mononuclear cell is induced into a DC cell; suspending lymphocyte and the wall-attaching DC cell are separated; a tumor antigen peptide is selected to conduct antigen supporting on the DC cell so as to obtain a mature DC cell; the suspending lymphocyte is cultured to obtain a CIK cell; the mature DC cell is added to the CIK cell to perform large-scale co-culture, then a DC-CIK cell is collected, detection is performed, the cell is put in an immune cell bank for cryopreservation after being qualified, and a cell file is established; a corresponding effect cell type is selected from the immune cell bank according to clinical diagnosis, and retransformation is performed after resuscitaion culture for 24 hours. The immune cell bank established by adopting the method is a set of various tumor antigen peptide effect cells, patients can select appropriate effect cells for resuscitaion and retransformation when needing cell treatment, the time waiting for cell culture is shortened, the best treatment period is mastered, and a treatment effect is improved.

The invention belongs to the field of biological medicine and specifically relates to a neural stem cell preparation for treating a Parkinson's disease through nasal delivery as well as a preparation method and application thereof. The preparation method comprises the following steps: cultivation and purification of seed cells; cultivation of P6-P8 generation cells; cultivation of a working cell repository; preparation of a preparation for treating the Parkinson's disease. According to the preparation as well as the preparation method and the application thereof, after the preparation is dropped or injected into a nasal cavity, the preparation can quickly form a solid membrane shape under a body temperature condition and can be adsorbed on the nasal cavity for slow absorption, and the preparation cannot enter an oral cavity or lungs along with breathing, so that the preparation can be used by patients and doctors conveniently and fast; in addition, test data indicate that the preparation has better curative effect and safety compared with a traumatic corpus striatum delivery. The preparation can be used for treating the Parkinson's disease, and an immunosuppressive agent is not needed.

A characterized cell library for EDA tools includes one or more mathematical models for each cell, and one or more preconditioning functions (and / or inverse preconditioning functions) for each mathematical model. Each mathematical model represents a performance parameter (e.g., delay, power consumption, noise) or a preconditioned performance parameter of the cell. The preconditioning functions convert an operating parameter (e.g., input slew, output capacitance) associated with the performance parameter into a preconditioned input variable for the mathematical models. In doing so, the preconditioning functions allow for more accurate modeling of complex data relationships without increasing the complexity (e.g., order and number of coefficients) of the mathematical models. Also, because the cell library can be substantially similar to conventional polynomial-based cell libraries except for the inclusion of preconditioning functions, preconditioning does not significantly increase storage requirements and conventional EDA tools can be readily adapted to use the preconditioned cell library.