Abstract: Microscopy imaging that allow for multiple mRNAs, proteins and metabolites to be spatially resolved at a subcellular level provides valuable molecular information which is a crucial factor for understanding tissue heterogeneity as for example within the tumor micro environment. The current invention describes a method (SUMI-Seq) which combines the use of Spatial Unique Molecular Identifier in situ sequencing and in vitro sequencing of rolonies derived from rolling circle amplification from padlock oligonucleotides targeting portion of RNA or cDNA transcript at a subcellular level with less limitation in the amount of transcripts and the length of the sequence that can be analyzed. Apart from padlocks oligonucleotides, the SUMI-Seq method can also be applied using circular oligonucleotides to spatially resolve proteins and metabolites to provide multiomics results.

Abstract: The present invention provides the use of a nucleic acid encoding SOCS1 for enhancing the efficacy of introducing at least one nucleic acid of interest into a cell; a method of repeated transfection of a cell with at least one nucleic acid of interest comprising the steps of adding a) nucleic acid encoding SOCS1, and simultaneously or subsequently b) at least one nucleic acid of interest encoding at least one polypeptide of interest, wherein at least step b) is repeated at least once; and a method of electroporation of a cell with at least one nucleic acid of interest comprising the steps of adding to the cell a) a nucleic acid encoding SOCS1, and simultaneously or subsequently b) said at least one nucleic of interest. The at least one nucleic acid of interest and the nucleic acid encoding SOCS1 may be mRNAs, wherein each of said mRNAs has a poly(A) tail at its 3? end comprising at least 200 adenines.

Abstract: A method to verify if a vibrational event has a higher intensity than a predefined intensity threshold, which is linked to a predefined frequency, includes: acquiring data detected by at least one accelerometer along respective X and Y axes, which are perpendicular to each other, determining values of frequency and intensity of at least one portion of the signals, derived from the acceleration data thus acquired along the two axes, when they exceed a threshold of predefined intensity, identifying, on the basis of the frequency determined, a predefined intensity threshold, and checking whether the intensity determined is higher or lower than the predefined intensity threshold identified, to establish whether or not the vibrational event from which the signals were derived has an intensity greater than the threshold of predefined intensity thus identified.

Abstract: The present invention provides a cell culture for obtaining an epithelial organoid, the cell culture comprising i) epithelial stem cells, or tissue fragments comprising said epithelial stem cells, ii) a basal medium for animal or human cells, iii) a Bone Morphogenetic Protein (BMP) inhibitor, iv) a mitogenic growth factor selected from the group consisting of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and keratinocyte growth factor (KGF), v) at least one soluble culture enhancer, wherein said at least one culture enhancer induces correct polarization of the cells in said cell culture within the developing organoid such as a laminin/entactin complex or entactin, and vi) a Wnt agonist if said epithelial stem cells, or tissue fragments comprising said epithelial stem cells are healthy cells, wherein said at least one soluble culture enhancer in said cell culture is a laminin/entactin complex in a con

Abstract: The present invention is directed to ligand like a chimeric antigen receptor (CAR), comprising an antigen binding domain specific for one or more antigens selected from the group consisting of CLA, CD142, CD73, CD49c, CD66c, CD104, CD318 and TSPAN8; cell populations expressing such CARs and the use of the cell populations for cancer therapy.

Abstract: The invention is directed to a method to provide a polynucleotide molecule comprising a first and a second strand with a barcode nucleotide sequence characterized in that the first strand is provided at its 5? end with an overhang of at least one universal base and the corresponding recessed 3? end of the second strand of the polynucleotide with at least one nucleotide provided with a blocking group, wherein the blocking groups are removed from the incorporated nucleotides by irradiation with light.

Abstract: The present invention provides an in-vitro method for analyzing a cell composition comprising human floorplate mesDA progenitor cells, the method comprising a) contacting the cells of said cell composition or the cells of a sample thereof with antigen binding molecules specific for the antigens FOXA2, OTX2, PAX6, and NKX6.1, thereby labeling the cells of said cell composition or of said sample, b) determining the percentage of said cells that are labelled with said antigen binding molecules for each of said antigens, and wherein the cells of said cell composition qualify as human floorplate mesDA progenitor cells if the protein expression profile of said cells is: 80-100% of said cells are positive for FOXA2, 80-100% of said cells are positive for OTX2, less than 10% of said cells are positive for PAX6, and less than 10% of said cells are positive for NKX6.1. A kit comprising said antigen binding molecules for use in said method is also provided.

Abstract: The present invention discloses an in vitro method for the generation of a cell composition comprising or consisting of ventral midbrain dopaminergic progenitor cells from a cell composition comprising pluripotent and/or multipotent stem cells, the method comprising the steps of A) differentiating said pluripotent and/or multipotent stem cells into ventral dopaminergic progenitor cells, thereby generating a cell composition comprising ventral dopaminergic progenitor cells comprising ventral midbrain dopaminergic progenitor cells and ventral hindbrain dopaminergic progenitor cells, and B) Enriching CD117 positive cells from said cell composition comprising ventral dopaminergic progenitor cells by using an antigen binding molecule specific for the CD117 antigen, thereby generating said cell composition comprising or consisting of ventral midbrain dopaminergic progenitor cells. Cell compositions obtainable by said method are also disclosed.

Abstract: The present invention is directed to a chimeric antigen receptor (CAR), comprising an antigen binding domain specific for MSLN in combination with one or more antigen binding domains specific for an antigen selected from the group consisting of CLA, CD66c, TSPAN8 and CD318; cell populations expressing such CARs and the use of the cell populations for cancer therapy.

Abstract: Microscopy imaging that allows for multiple mRNAs, proteins and metabolites to be spatially resolved at a subcellular level provides valuable molecular information which is a crucial factor for understanding tissue heterogeneity as for example within the tumor micro environment. The current invention describes a method (High Density-SUMI-Seq) which combines the use of Spatial Unique Molecular Identifier in situ localization and identification (by in situ sequencing or sequential fluorescence hybridization) of rolonies derived from rolling circle amplification of circular oligonucleotides and in vitro sequencing of target amplified RNA or DNA in combination with SUMI identification at a subcellular level with no optical diffraction limitation in the amount of amplified target information that can be analyzed per cell. Apart from amplified RNA or DNA, the High Density-SUMI-Seq method can also be applied using linear oligonucleotides to spatially resolve proteins and metabolites to provide multiomics results.

Abstract: The invention refers to a joint for hydraulic connection, including: an upstream body; a downstream body; and a joining device with screw, extended along a hinging axis, which crosses in succession the upstream body and the downstream body rotatably coupling them with respect to said hinging axis; being further provided an adhesive interposed between thread and counter-thread of the joining device.

Abstract: The invention is directed to a conjugate having the general formula (I): Xn—P—YmBo (I), with X is an detection moiety, P is a spacer unit, Y an antigen recognizing moiety, B an oligonucleotide comprising 2 to 300 nucleotide residues and n, m, o are independent integers between 1 and 100 wherein P and B are covalently bound to Y and X is covalently bound to P and wherein X is erasable. Further, the invention is directed to a library of such conjugates and a method of detecting target cells utilizing the conjugates or the library of conjugates.

Abstract: Microscopy imaging that allow for multiple mRNAs, proteins and metabolites to be spatially resolved at a subcellular level provides valuable molecular information which is a crucial factor for understanding tissue heterogeneity as for example within the tumor micro environment. The current invention describes a method (High Density—SUMI-Seq) which combines the use of Spatial Unique Molecular Identifier in situ localization and identification (by in situ sequencing or sequential fluorescence hybridization) of rolonies derived from rolling circle amplification of circular oligonucleotides and in vitro sequencing of target captured RNA or DNA in combination with SUMI identification at a subcellular level with no optical diffraction limitation in the amount of captured target information that can be analyzed per cell. Apart from captured RNA or DNA, the High Density—SUMI-Seq method can also be applied using linear oligonucleotides to spatially resolve proteins and metabolites to provide multiomics results.

Abstract: The present invention provides a combination comprising a) an antigen binding domain specific for CD90, and b) an antigen binding domain specific for CD326, for use in treatment of human cancer comprising cancerous cells that co-express CD90 and CD326. In one embodiment of the invention the combination comprises a) an immune cell comprising a CAR comprising an antigen binding domain specific for a tag of a first and a second polypeptide, b) said tagged first polypeptide that has an antigen binding domain specific for CD90, and c) said tagged second polypeptide that has an antigen binding domain specific for CD326, wherein the tag of the first polypeptide and the tag of the second polypeptide are identical. In a further embodiment the concentrations used for said first and that second polypeptide are below the activation threshold of said CAR, respectively, but the sum of both concentrations is above the activation threshold of said CAR.

Abstract: The invention is directed to a slide chamber comprising a top member (10), a fluidic seal (20), a transparent closure member (30) and base member (40) wherein the top member (10) is provided with at least one examination chamber (11) and a plurality of openings (12) positioned at two sides of the cover member outside of the examination chamber (11); and the base member (40) is provided with interconnecting means (41) complementary to the openings (12) of the top member characterized in that the interconnecting means (41) of the base member (40) and the openings (12) of the top member (10) are configured to mechanically interlock with each other by lateral movement thereby pressing the top member (10) against the transparent closure (30) member in a water-tight manner.

Abstract: The invention is directed to a process for extracting target cells from a three dimensional biological specimen by the steps imaging the three dimensional specimen; identifying target cells; registering the spatial parameters (x,y,z coordinates) of the target cells; and extraction of target cells according to their spatial parameters

Abstract: The invention is directed to a process for providing a cell comprising a chimeric antigen receptor (CAR) specific for one or more target antigens exposed on tumor microenvironment cells characterized by providing a cell sample comprising tumor microenvironment cells and non-tumor microenvironment cells and repeating the steps of—contacting the cell tissue with a conjugate comprising a fluorescent moiety and an antigen recognizing moiety—removing unbound conjugate from the cell tissue and detecting cells bound to the conjugate by the fluorescence radiation emitted by the fluorescent moieties of the first conjugates—erasing the fluorescence emitted by the fluorescent moieties of the conjugates until identifying at least two conjugates provided with antigen recognizing moieties recognizing different antigens, allowing in combination to discriminate between tumor microenvironment cells and non-tumor microenvironment cells and providing cells with the identified at least two antigen recognizing moieties as chimeri

Abstract: The invention is directed to a method to obtain the spatial location and sequence information of at least a part of a RNA or cDNA strand (006) in a sample comprising the steps a. hybridizing a first detection probe oligonucleotide (204) comprising 50-1000 nucleotides with its 3? and/or 5? end to the complementary part of the at least one RNA or cDNA strand, wherein the detection probe oligonucleotide is partially hybridized to a bridge oligonucleotide (205) comprising 5-100 nucleotides wherein a gap region (206) capable of binding oligonucleotides is created b. filling the gap region (206) in part with 1 to 16 barcode oligonucleotides comprising 4-20 nucleotides, wherein the barcode oligonucleotides determine the spatial information of the RNA or cDNA strand in the sample c.

Abstract: The present invention is directed to a method for identifying nucleic acids of a target cell from a cell population comprising—isolating at least one target cell from the cell population and at least one color-coded composition comprising a solid particle conjugated to an oligonucleotide into one compartment—lysing the isolated target cells—coupling the nucleic acid molecules of the lysed isolated target cells with the oligonucleotide of the color-coded composition forming a first conjugate—determining the sequence of the first conjugate, thereby identifying the target cell. characterized in that at least one target cell and the at least one color-coded composition are selected to be isolated into one compartment according to at least one pre-selected physical property of the target cell combined with at least one pre-selected physical property of the color-coded composition.

Abstract: The invention is directed to a Method for detecting differentiated hematopoietic cells comprising the steps: a) isolation of undifferentiated hematopoietic stem cells in groups of 1-1000 cells on a support b) proliferating the isolated cells to form cell colonies of differentiated hematopoietic cells by providing cell media comprising a growth factor c) contacting the cell colonies with one or more marker conjugates comprising at least one detection moiety and at least one antigen recognizing moiety against CD14, CD235a and CD15 d) detecting the relative amount of differentiated hematopoietic stem cells in a cell colony labelled with the marker conjugates.