Abstract: A method is disclosed for the in vitro growth and proliferation of germ cells obtained from the ovarioles of an insect. By culturing these cells in a medium supplemented with soluble cytokines and mitogenic agents and independent of feeder-cells, they can be induced to proliferate. Cells are stored by cryogenic means for future use.

Abstract: Disclosed is a novel biological sample preparation (BSP) device and a method of using such a device that enables inexpensive and flexible high throughput sample preparation and visualization for microscopy, including high resolution, single or multi-label, 2-dimensional (2D) or 3-dimensional (3D) fluorescence microscopic observation of biological samples, such as cultured cells. Also included in the invention are a high throughput chemical sample preparation device, and a method for to clone and screen cells using a BSP device.

Abstract: A cytological fixative and dehydrating agent used for treating histological and cytological tissues for pathological diagnoses consists of a blend of from about 11 to 21% ethyl alcohol, from about 35 to 45% methyl alcohol and from about 40 to 50% by weight isopropyl alcohol. The treated tissue is subjected to further processing to render the cells in the tissue stable prior to placing the tissue upon a glass slide for pathological examination.

Abstract: A method for producing a neuroblast and a cellular composition comprising an enriched population of neuroblast cells is provided. Also disclosed are methods for identifying compositions which affect neuroblasts and for treating a subject with a neuronal disorder, and a culture system for the production and maintenance of neuroblasts.

Abstract: This invention provides a method of assaying leukocyte binding to vascular tissue which comprises contacting a suspension of a monocyte-like cell line in a suitable medium with a human vascular tissue sample at a temperature of at least 10.degree. C., and quantitating the number of bound monocyte-like cells over a defined area of tissue sample. The assay allows agents which inhibit binding of monocytes to human vascular tissue to be identified and the invention also relates to the use of such agents in the therapy of atherosclerosis.

Abstract: A method of staining preselected, mounted specimens of either biological or nonbiological material enclosed within a staining chamber where the liquid staining reagents are applied and removed from the staining chamber using hypergravity as the propelling force. In the preferred embodiment, a spacecraft-operated centrifuge and method of diagnosing biological specimens while in orbit, characterized by hermetically sealing a shell assembly. The assembly contains slide stain apparatus with computer control therefor, the operative effect of which is to overcome microgravity, for example on board an International Space Station.

Abstract: A system for automatically detecting the presence of contaminants in samples. The system includes a microscope, controllable stage positioner for holding slides under the microscope, a computer for controlling the stage positioner and a digital camera to capture images through the microscope. The system scans microscope views of regions of a slide sample and provides the digital images to the computer. Image processing routines stored in the computer analyze the digital images and determine whether these images may contain certain contaminants by comparing the characteristics of the objects in the image with the known characteristics of the contaminants.

Abstract: Improved methods for the diagnosis and treatment of cancer which involve the targeting of slow-growing, relatively mutationally-spared cancer stemline are provided. These methods are an improvement over previous cancer detection and therapeutic methods because they provide for very early cancer detection and treatment and reduce the likelihood of clinical relapse after treatment.

Abstract: A method of treating a wound of a mammalian subject in need of such treatment, to promote healing thereof, comprising administering to the subject, e.g., to the wound locus, a composition comprising a fibroblast and keratinocyte proliferatingly effective amount of an amphipathic peptide, preferably an amphipathic peptide which is antimicrobially effective at such locus. A method is also disclosed of stimulating the accelerated growth of dermal tissue in a tissue culture containing same, comprising applying to the tissue culture a fibroblast and keratinocyte proliferatingly effective amount of an amphipathic peptide, by which the dermal tissue may be grown to produce skin for skin grafting purposes, utilizing a dermal tissue culture containing dermal tissue material of a skin graft recipient of such skin. Novel amphipathic peptides suitable for use in such methods are disclosed.

Abstract: A method for stimulating chondrocyte proliferation and inhibiting chondrocyte differentiation along the endochondral developmental pathway is provided comprising contacting condrocytes with an effective amount of Platelet-Derived Growth Factor (PDGF) such as PDGF-BB, PDGF-AA OR PDGF-AB in the substantial absence of growth factors which promote cell differentiation. This allows such cells to be multiplied in culture for loading onto a scaffolding material and implanting into a cartilage or bone wound.

Abstract: Methods are provided to prepare a variety of tissues in a pathology-stable form keyed to the use of an aqueous solution of C.sub.2-6 dialdehyde and/or dialdehyde addition products, including the group consisting of glyoxal and/or glutaraldehyde in an amount sufficient to prevent autolysis and other degradative changes in various tissues. The said solution is capable of treating a tissue so as to maintain the tissue in a condition suitable for pathology and/or other experimental observation. The stabilizing solution is also useful to prepare parts or whole animals and plants in an anatomically preserved state for a prolonged period of time, and compositions useful in that method.

Abstract: The activation of neurons in specific areas of a mammalian brain are determined by measurement of levels of mRNA following administration of an agent that potentially affects neuronal activity. Differential levels of mRNA caused by an agent can be measured, as can the specific distribution of the alteration of neuronal activity. Many mental disorders are associated with the abnormal function of a particular area of the brain. The present invention allows the determination of which area of the brain, if any, is affected by an administered agent. Accordingly, the invention provides a valuable tool for assaying compounds as potential therapeutic agents.

Abstract: The invention discloses a process for sorting and harvesting biological objects on a planar carrier (2). On a planar carrier (2) an object field or the object itself, located on the carrier (2), is cut out with a laser beam (6) and transferred by means of a laser-induced transport process to a collector substrate (5) which is disposed directly above or below the carrier. During the cutting-out process, either the laser beam (6) moves in a closed curve about the object or the object itself is cut directly out of the carrier (2) in a computerized manner. This method enables individually selected objects to be spatially separated and sorted from a very large number of objects. The method can also be used to separate specific cells from tissue sections.

Abstract: The invention relates to an optical system for determining the distribution, environment, or activity of fluorescently labeled reporter molecules in cells for the purpose of screening large numbers of compounds for specific biological activity. The invention involves providing cells containing fluorescent reporter molecules in an array of locations and scanning numerous cells in each location with a fluorescent microscope, converting the optical information into digital data, and utilizing the digital data to determine the distribution, environment or activity of the fluorescently labeled reporter molecules in the cells. The array of locations may be an industry standard 96 well or 384 well microtiter plate or a microplate which is a microplate having a cells in a micropaterned array of locations. The invention includes apparatus and computerized method for processing, displaying and storing the data.

Abstract: The present invention provides a cell culture medium useful for a biochemical analysis of antioxidant function in human lymphocytes, said medium comprising, a buffered, serum-free solution containing the following ingredients: a carbohydrate selected from the group consisting of glucose and a compound biologically capable of producing glucose in the cells, a biologically usable form of pantothenic acid, choline or a biological usable form of a substance capable of producing choline in the cells, inorganic ions comprising chloride, phosphate, calcium, magnesium, potassium, sodium, and iron in a biologically utilizable form, cumene hydroperoxide, deionized water, and a mitogen in an amount effective to stimulate the lymphocytes being assayed; said buffered, serum-free solution having a pH from about 6.8 to 7.6, said cell culture medium characterized by being effective to determine nutritional deficiencies, inadequacies, and imbalances and to biochemically analyze antioxidant function of the lymphocytes.

Abstract: Systems and methods for acquiring laser capture microdissection samples are disclosed. A method of making a laser capture microdissection consumable includes providing a transfer film carrier having a substrate surface; and fabricating a laser capture microdissection transfer film on said substrate surface, wherein forming includes hot vacuum baking said laser capture microdissection transfer film. The systems and methods facilitate quick and accurate laser capture microdissection while simultaneously minimizing contamination.

Abstract: The present invention provides a method of treating cancer comprising (a) obtaining a tumor cell line, (b) modifying the tumor cell line to render it capable of producing an increased level of a cytokine relative to the unmodified tumor cell line, and (c) administering the tumor cell line to a mammalian host having at least one tumor that is the same type of tumor as that from which the tumor cell line was obtained, wherein the tumor cell line is allogeneic and is not MHC-matched to the host. The present invention also provides a pancreatic tumor cell line, a method and medium for obtaining such a tumor cell line, and a composition comprised of cells of a purified pancreatic tumor cell line.

Abstract: A dual purpose tissue fixative is described. The fixative comprises a cross-linking aldehyde, alcohol, a chelating agent and a non-amine buffer. More particularly, the tissue fixative comprises 0.1-2% w/v formaldehyde, 45-90% w/w alcohol, 1-10 mM of a chelating agent and 1-50 mM of a non-nitrogen buffer. The tissue fixative permits the recovery of high molecular weight DNA and RNA from the tissue sample for molecular genetic analysis. The tissue fixative also preserves the morphology and immunogenicity of the tissue allowing for pathology analysis. The tissue fixative is compared to 95% alcohol and a standard fixative as 10% BNF.

Abstract: Stable solid forms of preservative and embalming compositions are described which can be prepared by adding a sufficient amount of a chemical or chemicals containing at least one polar group capable of forming hydrogen and Van der Waal bonds with the reactive moieties of the aldehydes and antimicrobial agents in the preservative and embalming compositions. The amount of polar chemical(s) necessary to stabilize the reactive aldehydes and antimicrobial compounds/agents is based on a molar ratio of (i) polar groups in the stabilizing chemicals to (ii) reactive groups in the active preservative ingredient of at least 0.8.

Abstract: The invention provides a cell control product for use with a cytoenzymology assay to confirm enzymatic activity in devices employing electronic and/or optical means. The cell control product comprises a lyophilized mammalian cell which is capable of being rehydrated in water to exhibit cellular structure and cellular enzymatic activity so that said lyophilized cells can function effectively as a cytoenzymology cell control in an enzymatic analysis. The cellular structure of the cell control is capable of being authenticated by light scatter or microscopy analysis and the cellular enzymatic activity is capable of being authenticated by fluorescence analysis. The cell control exhibits a real time stability when stored at between 2 and 8.degree. C. for at least a two month period of time. Preferably the mammalian cell is an abnormal cell selected from Molt 4, CCRF-CEM and HL-60 cell lines.

Abstract: A method for detecting the presence of a double-stranded target nucleic acid sequence contained in fixed cells or cell structures is provided. The method includes the steps of forming the fixed cells or cell structures by fixing cells or cell structures so as to allow a nucleic acid probe to enter, and forming a probe/RecA complex in which a single-stranded probe and RecA protein are stably bound to each other. The probe/RecA complex is allowed to react with the double-stranded target nucleic acid sequence to bind thereto under conditions in which the double-stranded target nucleic acid sequence is not denatured, and by detecting the RecA protein included in the probe/RecA complex, the presence of the double-stranded target nucleic acid sequence is detected.The invention provides a diagnostic method by which the position of a specific gene or its regulatory region in a chromosome and the presence of a nucleic acid sequence derived from virus can be measured or visualized with high sensitivity and with ease.

Abstract: The present invention relates to a method of stimulating the proliferation and appropriate cell maturation of a variety of different cells and tissues in three-dimensional cultures in vitro using TGF-.beta. in the culture medium. In accordance with the invention, stromal cells, including, but not limited to, chondrocytes, chondrocyte-progenitors, fibroblasts, fibroblast-like cells, umbilical cord cells or bone marrow cells from umbilical cord blood are inoculated and grown on a three-dimensional framework in the presence of TGF-.beta.. Stromal cells may also include other cells found in loose connective tissue such as endothelial cells, macrophages/monocytes, adipocytes, pericytes, reticular cells found in bone marrow stroma, etc.

Abstract: An in vitro method of identifying or isolating fetal cells from a blood sample is described. Fetal nucleated erythrocytes or erythroblasts are identified by using an antibody or antibody fragment specific for embryonic hemoglobin or an embryonic hemoglobin chain. Once the fetal cells are identified, they can be treated to render the fetal nucleic acids or proteins available for identification or amplification. Detecting the occurrence or existence of selected fetal nucleic acids or proteins allows a quantitative or qualitative diagnostic or prenatal evaluation, including determining the sex of the fetus, determining chromosomal, single gene or protein abnormalities, and determining the presence or absence of particular genes, nucleic acid sequences or proteins.

Abstract: The invention features a method for the selection and expansion of bone marrow stromal cells. The method includes the steps of obtaining bone marrow stromal cells; introducing the stromal cells into a vessel pre-coated on an inner surface with a gelatin, and containing a culture medium including an acidic fibroblast growth factor ("aFGF") polypeptide; and expanding the stromal cells in the culture medium under conditions and for a time sufficient to obtain an increased number of bone marrow stromal cells. The culture medium additionally can include heparin, and the vessel additionally can be precoated with fetal bovine serum.

Abstract: A method is disclosed for gene transfer into a culture of primitive stem cells which comprises a prestimulation step of adding a blocking agent to block at least one inhibitor of a cell cycle of said primitive stem cells. The prestimulation is time-limited for a period of less than approximately 36 hours so that said culture of primitive stem cells retains hematopoietic potential.

Abstract: Disclosed is a support retaining member (216) for use in processing a sample supported on a support (6), the member (216) being such than when assembled together with a support (6) it forms a support cell, the support cell comprising a substantially sealed chamber, the chamber being provided with a fluid inlet and a fluid outlet for the introduction and removal respectively of fluids used in processing the sample.

Abstract: The present invention relates to a DNA construct containing a cDNA sequence for human poly(ADP-ribose)-polymerase, cell lines containing the DNA construct, and a process for identifying DNA-damaging substances by means of these cell lines which overexpress poly(ADP-ribose)-polymerase.

Abstract: A method is disclosed for producing magnetic or piezoelectric particles consisting of dried microorganisms which contain fixed magnetic or piezoelectric powder. The microorganisms are cultured in a nutrient medium containing the magnetic or piezoelectric powder which is then fixed or ingested by the microorganism. The microorganisms containing the magnetic or piezoelectric powder are then extracted from the nutrient medium and dehydrated to obtain dry particles. The dry particles may be compacted or inserted into a matrix to form a composite material.

Abstract: A method and single solution denaturation and hybridization solution useful with synthetic oligonucleotide probes for hybridizing DNA or RNA sequences by in situ hybridization. The inventive protocol employs synthetic oligonucleotide probes and a glycerol-based hybridization solution which is particularly useful in fluorescence labeling of chromosomes X, Y, 13, 18 and 21 in human lymphocytes, amniocytes and metaphase chromosomes, with the label being sufficiently stable to permit archival storage of the labeled sample for later hybridization or analysis.

Abstract: An article and method to create retrievable sites of implantable and transplantable bone marrow, in a human or animal recipient, by implanting bone-marrow inducing conjugates of collagen and demineralized freeze-dried bone allograft (DFDBA) into accessible soft or hard tissue sites is provided. The implantation of such collagen-DFDBA conjugate grafts into suitable ectotrophic sites results in the formation of bone-marrow and hematopoietic tissue within the implant. By removing all or a portion of the bone marrow containing areas of the implant, an additional source of hematopoietic tissue and bone marrow cells is formed which can subsequently be transplanted into the same individual as an autograft, or a related or compatible individual as an allograft.

Abstract: A serum-free medium which supports the proliferation and differentiation of CD34.sup.+ cells purified from normal bone marrow, peripheral blood of patients treated with cytokines, and umbilical cord blood is described. The recipe for the formulation is given, which provides a medium suitable for the proliferation and differentiation of CD34.sup.+ cells for use in human therapeutic protocols.

Abstract: A composition and method for staining cellular DNA are disclosed. The composition of the present invention is an aqueous alcoholic solution that includes a cationic stain, a metabisulfite, and an alcohol preferably selected from methanol, ethanol, and mixtures thereof. In a preferred embodiment, the cationic stain is thionin.

Abstract: The present invention provides methods for rapidly determining the proportion of live, dead and stressed cells in a given microbial culture. The measurement of stressed cells is used as a quantitative indicator of the relative health of the population and the population's ability to withstand long term stress. The present methods comprise the steps of(i) staining the culture with a combination of membrane-permeable and membrane-impermeable stains; and(ii) quantifying the population of stressed cells, using the measurement thus obtained as an indicator of the relative health of the population.

Abstract: Assays and kits for identifying fertile sperm samples (e.g. for use in an assisted reproductive technology or as an indication of an ineffective male contraception (vasectomy)) and sub-fertile sperm samples (e.g. as an indication of a potentially infertile male donor or an effective male contraception) are disclosed.

Abstract: For the fixated cryopreservation of single living biological objects or s objects compiled in a given number (for example cells) a cryogenically cooled substrate (13) is jetted therewith in a enveloping solution in microdroplet form (12) from a storage vessel, for example by means of a microdroplet jetting device (11). The substrate is temperature-controlled via a coolant (15), the surface to be jetted being located in a gas atmosphere or in vacuum (14). The substrate surface is maintained at a temperature T1 resulting in freezing of the impinging microdroplet, the substrate surface being possibly supportingly microstructured and comprising sensing elements. By controlled movement of either the substrate or the microdroplet jetting device the microdroplets can be applied singly and in patterns in arrays freely selectable or predetermined by the structuring of the substrate.

Abstract: Provided is a medium for culturing normal human epidermal melanocytes in vitro. The medium comprises a basal medium for culturing animal cells, Ca.sup.2+ at a final concentration of between about 0.15 mM and about 1.2 mM and Mg.sup.2+ at a final concentration of 1.2 mM and 6 mM or Ca.sup.2+ at a final concentration of between about 0.9 mM and about 1.2 mM and Mg.sup.2+ at a final concentration between about 0.6 mM and 6.0 mM and 0.001 to 0.1% scrum (v/v). The medium can promote both dendrite formation and proliferation of the cells.

Abstract: Media and methods are disclosed for the in vitro formation of a histologically complete human epithelium. The media are serum-free, companion cell or feeder layer free and organotypic, matrix free solutions for the isolation and cultivation of clonally competent basal epithelial cells. The media and methods of the invention are useful in the production of epithelial tissues such as epidermis, cornea, gingiva and ureter.

Abstract: Haematopoietic stem cells (which term includes early progenitor cells) are immobilized on a substrate coated with a fibrin matrix and including a substance capable of both binding to the fibrin matrix and also having an RGD amino acid sequence for binding to the stem cells. The substance may be fibronectin or thrombospondin. The substrate is generally in the form of a closed bag formed of a carbon dioxide-permeable and oxygen-permeable plastics material which allows culturing of the stem cells. The cultured stem cells may re-engraft a patient following chemotherapy or to correct haemotological deficiencies. Stem cells may be harvested from peripheral blood onto the coated substrate. The stem cells in contact with the coated substrate are good candidates for gene therapy to introduce a heterologous gene e.g. employing a transfection vector.

Abstract: The present invention relates to a culture of human olfactory neurons. The neurons may display a normal neuronal pathology or a pathology characteristic of a generalized central nervous system disease. The cultured neurons can be used for neurotoxicity tests, screening for therapeutic drugs and anti-viral agents.

Abstract: A composition and method for maintaining the viability of human mesenchymal precursor cells in a serum-free environment which composition includes (1) a minimum essential medium; (2) serum albumin; (3) an iron source; (4) insulin or an insulin-like growth factor; and (5) at least one amino acid selected from the group consisting of glutamine, arginine and and cysteine, and is free of serum. Also, a composition and method for culture expanding human mesenchymal precursor cells in a serum-free environment. This composition further includes a mitogen, paricularly a serotonergic agonist. The cells are preferably isolated human mesenchymal stem cells.

Abstract: Disclosed are a composition of chemically defined components which support the in vitro chondrogenesis of mesenchymal progenitor cells, a method for in vitro chondrogenic induction of such progenitor cells and a method of forming human chondrocytes in vitro from such progenitor cells.

Abstract: A method for the rapid diagnosis of aerobic bacteria present in a biological medium comprising successively:a) a rapid culture under aerobic conditions of a sample of the biological medium, with stirring,b) a rapid centrifugation in order to recover the bacteria from the centrifugation pellet,c) the projection, by centrifugation, of the pellet thus obtained onto a supportd) the revealing of bacteria on the support.

Abstract: A method for diagnosis or prognosis of a cancer is disclosed. The method comprises (i) detecting in a first biological sample protein tyrosine phosphate .alpha. (PTP.alpha.) or PTP.alpha. nucleic acid, and (ii) comparing the level of PTP.alpha. or PTP.alpha. nucleic acid in the first sample with the level in a second biological sample known to be from normal tissue. Any overexpression of PTP.alpha. or PTP.alpha. nucleic acid in the first sample compared to the second sample is indicative that the first sample is from a cancerous tissue.

Abstract: A method of inducing the proliferation and/or differentiation of human adult pancreatic cells entails contacting primary cultures of such cells with Hepatocyte Growth Factor/Scatter Factor (HGF/SF), thereby inducing a proliferation of .beta.-epithelial cells, an increase in the number of .beta.-epithelial cells which form islet-like cell clusters, and an increase in insulin production per cell. The method is improved by culturing the cells on an extracellular matrix such as 804G in the presence of HGF/SF, and is further improved by reaggregating thus-treated cells and contacting said cells with an insulin gene upregulating agent such as a poly(ADP-ribose) synthetase inhibitor such as a nicotinamide or benzamide. The method provides increased numbers of functional islet-like cell clusters for transplantation.

Abstract: Immortalized human stromal cell lines sustain and expand human hematopoietic precursor cells. The precursor cells are obtained from a blood product and inoculated into a culture medium conditioned by exposure to a human stromal cell line. Preferred human stromal cell lines secrete SCF, LIF, MIP1.alpha., and IL-6, as exemplified by a human stromal cell line designated HS-1. The conditioned culture medium may be supplemented with additional growth factors, such as interleukin-3. After expansion the human hematopoietic precursor cells are harvested and returned to a patient or frozen and stored. The immortalized human stromal cell lines can also be used as feeder layers in ex vivo bone marrow cultures or in colony forming assays.

Abstract: Method for treatment of a disease in a patient characterized by accumulation of .beta.-amyloid. The method includes identifying a patient potentially suffering from such a disease and contacting a neuron of the patient with a therapeutically effective amount of a tachykinin agonist. Methods for screening for compounds useful for treating such a disease are also disclosed.

Abstract: The invented methods are effective for the detection and quantification of bacterial spores in a sample medium. A lanthanide such as europium or terbium is combined with a medium to be tested for endospore content. The lanthanide will react with calcium dipicolinate present in any bacterial spores in the sample medium to produce a lanthanide chelate, specifically, terbium or europium dipicolinate. The lanthanide chelate has distinctive absorbance and emission spectrums that can be detected using photoluminescence testing, for example. The occurrence of emission from the sample medium upon excitation at wavelengths distinctive of the lanthanide chelate thus reveals the presence of spores in the sample medium. The concentration of spores can be determined by preparing a calibration curve that relates absorbance or emission intensities to spore concentrations for test samples with known spore concentrations.

Abstract: A sensor system and method are provided that are capable of the real-time detection of target live microorganisms, such as biological warfare agents. The sensor system includes a highly-sensitive, highly-selective sensor cell that comprises a single-stranded oligonucleic acid sequence that is complementary to a portion of the DNA of a target live microorganism, the oligonucleic acid having been modified with the covalent attachment of electron donor and acceptor moieties. In the presence of the targeted microorganism, hybridization occurs between the modified oligonucleic acid and the microorganism's DNA, such that the electron conductance between the electron transfer moieties greatly increases, thereby providing a means of detecting the presence of the target live microorganism.

Abstract: Human pancreatic endocrine cells are proliferated without loss of hormone function in a culture medium containing extracellular matrix from bladder carcinoma cell lines in the substantial absence of hepatocyte growth factor/scatter factor. Proliferation is preferably carried out in the substantial absence of any peptide growth factors and nicotinamide. The cells may be proliferated in a monolayer on a solid substrate. Islets and islet-like cell clusters are proliferated without loss of insulin-secreting function by incubation in a medium containing extracellular matrix from a human bladder carcinoma cell line, preferably cell line ATCC HTB-9.

Abstract: A method for staining fecal specimens to diagnose stool parasites preserved with a non-mercury fixative wherein the preserved specimen is stained with a composition of iron hematoxylin stain and trichrome stain.