84 results about "Immunophenotyping" patented technology

Devices and methods for automated collection and image analysis are disclosed that enable identification or classification of microscopic objects aligned or deposited on surfaces. Such objects, e.g. detectably labeled rare target cells, are magnetically or non-magnetically immobilized and subjected to Time Delay Integration imaging (TDI). Incorporation of TDI technology into image cytometry analysis, like CellTracks®, makes it possible to image moving objects with very high sensitivity and signal-to-noise ratios. Implementation of TDI camera technology with dual excitation and multispectral imaging of enriched rare cells provides a rapid system for detection, enumeration, differentiation and characterization of imaged rare cells on the basis of size, morphology and immunophenotype.

The present invention provides an isolated population of adult hematopoietic stem cells (HSC) derived from embryonic stem cells (ESC) induced to differentiate in vitro by culturing ESCs in a medium with stem cell factor, interleukin (IL)-3, and IL-6. HSC of immunophenotype c-kit+ or c-kit+ / CD45+ from this population are isolated and injected either intra bone marrow or intravenously into myeloablated recipient individuals. This method allows for the establishment of banks of allogeneic ESC-derived adult stem cells for treatments of autoimmune diseases, immune deficiencies and induction of immunotolerance during organ transplantation. These allogeneic ESC-derived adult hematopoietic stem cells (HSC) may be used in reconstituting bone marrow, without the development of teratomas or graft versus host disease, despite crossing histocompatibility barriers. Additionally, allogeneic ESC-derived HSC can be used to prevent the development of autoimmune diseases or organ rejection during transplantation.

The invention discloses a cryopreservation solution for preserving congenital intrathoracic kidney (CIK) cells and application thereof. The cryopreservation solution for preserving the CIK cells is prepared according to the following methods: adding dimethyl sulfoxide and dextran into a lymphocyte serum-free medium, so that the volume percent content of the dimethyl sulfoxide is 5-15 percent, and the volume percent content of the dextran is 5-15 percent. Compared with the cells before cryopreservation, the cells cryopreserved by employing the cryopreservation solution have the following advantages: (1) the cellular morphology does not have obvious difference; (2) immunophenotype (CD3+CD56+CD8+) does not have obvious difference; and (3) the cell proliferation activity and cell killing activity do not have obvious difference.

The present invention encompasses methods and compositions for generating an isolated adipose tissue-derived stromal cell exhibiting a low level of immunogenicity. The present invention encompasses methods and compositions for reducing an immune response associated with transplantation by administering the recipient with an amount of adipose tissue-derived stromal cells effective to reduce or inhibit host rejection and / or host versus graft disease.

The invention provides a method for culturing human amniotic mesenchymal stem cells (hAMSCs) by enzyme digestion and EGF (epidermal growth factor). The method comprises the following steps of hAMSCs separation, hAMSCs primary culture, hAMSCs sub-culture and amplification, hAMSCs cryopreservation and recovery, and detection of hAMSCs cytometric immunophenotyping. According to the method, amniotic extracells are collected by digesting trypsin and collagenase for multiple times step by step, and adherent culture and digestion time control are combined by addition of 4ng / ml EGF, so that human amniotic mesenchymal stem cells (hAMSCs) with high purity and high activity can be obtained. The enzyme digestion and EGF adopted in the method have the advantages of simpleness in operation, quickness and practicality and the like, so that the method is capable of effectively separating and purifying hAMSCs.

The invention relates to an application of autologous adipose-derived mesenchymal stem cells in preparation of medicines for treating joint diseases. The application is characterized in that adipose-derived mesenchymal stem cells (Ad-MSCs) separated and extracted from autologous adipose tissues of the patient are injected to the diseased articular cavity in a large dose once. The adopted mesenchymal stem cells have immunophenotyping and representing biological marks of Ad-MSC through flow cytometry authentication, and can be directionally induced and differentiated to osteoblast and chondrocyte. Primary cells are directly injected to the articular cavity of the diseased joint to treat various joint diseases. With the adoption of the autologous adipose-derived mesenchymal stem cells as the primary cells, the cells have better differentiative potential and the ability of inhibiting inflammation and immunoregulation ability. The adipose-derived mesenchymal stem cell transplant provides a novel effective means for treating osteoarticular diseases. The method is simple and feasible, and the quantity of cells collected is great and the trauma is small. The autologous mesenchymal stem cells of the patient are applied without invading related ethics, so that the risk of immunological rejection, infectious diseases and the like is avoided.

The invention provides a reagent composition for leukemia / lymphoma typing. The reagent composition comprises 19 kinds of antibodies. According to the optimized antibody combination, the fluorescence labeling combination of the corresponding antibody and the result interpretation method, the AML, the ALL-B, the ALL-T, the MPAL, the NHL-B, the NHL-T, the PCN, the NHL-NK and the chronic myeloid tumors can be preliminarily screened comprehensively and efficiently only by using 19 antibodies and one-tube cell one-time sample loading, and the aim of clearly diagnosing the AML, the ALL-B, the ALL-T, the MPAL, the NHL-B, the NHL-T and the PCN is achieved. The 19 antibody combinations are used for analyzing the expression of tumor cell antigens in a sample, and leukemia-related immunophenotypes (LAIP) which can be used for monitoring post-treatment trace residual disease (MRD) in the antibody combinations are determined.

The invention discloses an acute B-lymphocyte leukemia initiated cell phenetic classification kit. The kit is used in cooperation with a sever-color flow cytometer. The structure of the kit is configured with the following fluorescently-labeled monoclonal antibody: CD58FITC, CD10PE, CD34PerCP, CD38APC, CD19APC-Cy7 and CD45PacificBlue. The kit disclosed by the invention can be used for fast and accurately detecting the immunophenotype of the leukemia initiated cell of a B-ALL patient in first visit; and the CD58 expression proportion on the leukemia initiated cell tested by the kit provided by the invention can be used as one of the prognostic indicators of the B-ALL patient, and the indicator has important guiding significance in the prognosis of the B-ALL patient and the decision of a clinic treatment scheme.

The invention belongs to the technical field of cellular immunology, in particular to a method for amplifying human peripheral blood gamma / deltaT cells massively and preferentially in vitro. The method comprises the following steps of: (1) preparing tubercle bacillus heat stable antigen by using the cultured tubercle bacillus; (2) stimulating to amplify the gamma / deltaT cells by combining the prepared tubercle bacillus heat stable antigen and cell factors rhIL-2 and rhIL-21; and (3) performing morphologic observation through a microscope, performing purity and immunophenotyping analysis by using a flow cytometry, and performing cytotoxicity detection through MTT assay. The method has the advantages that: culture conditions and operation are simple, a large number of gamma / deltaT cells with high purity and high cytotoxicity can be prepared in short time, and the cost is low.

The invention discloses a human umbilical mesenchymal stem cell and a preparation method thereof. The human umbilical mesenchymal stem cell is prepared by carrying out aseptic treatment, mechanical shearing, Wharton jelly exposure, collagenase digestion and isolated culture on an umbilical stalk of a healthy fetus by a term abdominal delivery. After freezing and thawing, the cell activity does not decline dramatically, thereby ensuring that the human umbilical mesenchymal stem cell is an ideal seed cell for cellular therapy. The invention also relates to a primitive mesenchymal stem cell prepared by the method, and the primitive mesenchymal stem cell is confirmed as the mesenchymal stem cell by the identification of morphology, immunophenotype and invitro differentiation experiments in accordance with the mesenchymal stem cell standards established by International Society for Cellular Therapy (ISCT). The cell count, cell activity, purity, doubling time and multiplication capacity of the human umbilical mesenchymal stem cell are apparently higher than those of bone marrow-derived mesenchymal stem cell.

The system includes a simple system for CD4 and CD8 counting in point-of-care HIV staging within resource poor countries. Unlike previous approaches, no sample preparation is required with the sample added directly to a chip containing dried reagents by capillary flow. A large area image cytometer consisting of an LED module is used to excite the fluorochromes PerCP and APC labeled targets and a monochrome CCD camera with a combination of two macro lenses captures images of 40 mm2 of blood (approximately 1 micro liter). CD4 and CD8-T-lymphocyte counts correlate well with those obtained by flow cytometry. The cytometer system described in the present invention provides an affordable and easy-to-use technique for use in remote locations.