Abstract: A cell is separated from a culture broth with an extremely simple configuration, and production yield of a target product is improved. There are provided separating means for separating a cell from the culture broth containing the cell, a buffer tank connected to the separating means through a first pipeline, at least one or more liquid reservoir tank connected to the buffer tank through a second pipeline, a first valve disposed on the first pipeline and controlling communication between the separating means and the buffer tank, and a second valve disposed on the second pipeline and controlling communication between the buffer tank and the liquid reservoir tank.

Abstract: The present invention has its object to provide a material for separating stem cell and a filter for separating stem cell, each is capable of selectively separating and recovering, in a simple and easy manner, stem cells from body fluids or biological tissue-derived treated fluids, a method for separating and recovering stem cells, and stem cells obtained by such method. The present invention is a material for separating stem cell which has a density K of 1.0×104?K?1.0×106 and a fiber diameter of 3 to 40 ?m; a filter for separating stem cell which comprises the material for separating stem cell as packed in a container having a fluid inlet port and a fluid outlet port; a method of separating and recovering stem cells which comprises using the material for separating stem cell or the filter for separating stem cell; and a method of producing a multipotent cell fraction.

Abstract: The present invention relates to a pharmaceutical composition comprising of preparations of human embryonic stem (hES) cells and their derivatives and methods for their transplantation into the human body, wherein transplantation results in the clinical reversal of symptoms, cure, stabilization or arrest of degeneration of a wide variety of presently incurable and terminal medical conditions, diseases and disorders. The invention further relates to novel processes of preparing novel stem cell lines which are free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor and conditioned media. This invention further relates to the isolation, culture, maintenance, expansion, differentiation, storage, and preservation of such stem cells.

Abstract: Disclosed is a hematopoietic stem cell proliferation inducing agent which comprises a pulverized product of placenta-constituting cells as an active ingredient and can induce/proliferate a hematopoietic stem cell in the peripheral blood. Since the proliferation inducing agent can induce/proliferate a hematopoietic stem cell to increase the amount of hematopoietic stem cells in the peripheral blood, it becomes possible to ensure the increase in the amount of hematopoietic stem cells in the peripheral blood. The proliferation inducing agent can be used for the prevention and treatment of various diseases which are caused or believed to be caused by the decrease in the amount of hematopoietic stem cells (e.g., leukemia, malignant lymphoma, aplastic anemia, Alzheimer's disease, Parkinson's disease, dilated cardiomyopathy, myocardial infarction) and produces extremely small adverse side effects.

Abstract: In one embodiment, the method comprises providing tissue, preparing the tissue, and treating the tissue to improve remodeling characteristics of the tissue. The tissue may be, for example, cortical bone. Treating the tissue to improve remodeling characteristics may comprise heating the tissue, treating the tissue with a chemical, or other. Heating the tissue may be done in the absence of oxygen and may comprise heating the tissue in a vacuum, heating the tissue in an inert atmosphere, heating the tissue in a reducing atmosphere, coating the tissue with a protective coating and heating the tissue, or other. Further embodiments comprise treating the tissue in supercritical fluids, for example, to dry or virally inactivate the tissue.

Abstract: A hybrid bioreactor for cell culture is disclosed. To simultaneously apply compressive strain for cell differentiation and shear strain for cell proliferation to cells, the hybrid bioreactor includes a plurality of reactor tube assemblies (100), a compressive strain motor (5), a shear strain motor (25), a lower anchor mount (20) having a plurality of toothed anchors (70) to respectively anchor the lower ends of the reactor tube assemblies (100) to the lower anchor mount (20), a ball screw (90) operated in conjunction with the compressive strain motor (5), an upper anchor mount (60) which engages with the ball screw (90) to vertically move upward and downward and having a plurality of compressive strain anchors (80) to anchor the upper ends of the reactor tube assemblies (100) to the upper anchor mount (60), a power transmission unit to transmit the rotating force of the shear strain motor (25) to the toothed anchors (70).

Abstract: The present invention provides a process and kit for isolating cone photoreceptor cells from retinal tissue with a purity level of at least 80%, typically of about 90%. The isolation process uses a PNA-panning procedure conducted on dissociated retinal tissue. The present invention also provides a culture medium enabling the in vitro survival and development of such isolated cone cells. The means of the invention are applicable to adult mammalian cone cells, and more particularly to adult human cone cells. They have the advantage of being applicable to pathologic or otherwise altered cone cells, and thus give access to the screening of compounds capable of showing neuroprotective activity on adult cone cells.

Abstract: There is provided a method for growing human intervertebral cells. Disc tissue is surgically removed from a normal disc of a patient, the cells expanded by feeding with a cell stimulant such as a growth factor, or a cytokine or a bioactive agent to form monolayer primary cell cultures on a plastic mesh such as a nylon mesh. In the case of a growth factor, fetal bovine serum is preferred as it improves cell proliferation and production of appropriate extracellular matrix components. In another aspect of this invention, the monolayer primary cell cultures are seeded in alginate or agarose and fed again with the cell stimulant until three-dimensional cell cultures are formed. The cells are recovered from the alginate or agarose or from monolayer cultures. Re-implantation is carried out using bioresorbable carriers or cell suspensions.

Abstract: A method of processing an organ is disclosed. The method comprises: (a) placing an organ in a sealable container; (b) disrupting the structure of said organ to yield a cell suspension; and (c) transferring said cell suspension to a sealable cell-suspension storage container, thereby isolating cells of said organ, wherein said sealable container, wherein said disrupting and said transferring are all performed substantially in a continuous vessel.

Abstract: An extract for preventing or treating thrombotic disease, particularly, an extract of leech and/or earthworm with molecular weight of not more than 5800 Dalton, and processes for preparation, pharmaceutical compositions and uses thereof. The extract of the present invention has improved significantly safety without any reducing in pharmaceutical activities or the therapeutical effects as compared to existing products.

Abstract: This invention relates to methods and apparatus for cryopreserving biological materials for extended periods of time. In an exemplary embodiment, the method comprises suspending biological materials in a cryosolution, freezing the biological materials in the apparatus, and removing the frozen biological materials from the apparatus to thaw them for use. In another embodiment, a cell cryopreservation solution is provided which includes 20% Dimethyl Sulfoxide (DMSO) to maintain the viability of cells upon freezing, storage, and thawing. The media can be used for cryopreservation of a wide variety of different cell types from various sources. In addition, an apparatus that facilitates the storage of cells and the subsequent removal of the cells for thawing to permit substantially direct inoculation of a bioreactor is disclosed.

Abstract: The invention relates to a method for cultivating cancer cells for scientific serial assays, wherein a tissue sample which is heterogeneous with respect to contaminants, normal cells and tumor cells is locally separated in a sequential-parallel splitting method. The locally separated sample segments are further split, wherein the tissue fragments and liquids of the tissue segments are separately placed in a given cell culture medium and grown under predetermined culture conditions. The invention also relates to a cell culture medium and a device for splitting the tissue samples into disc segments. The inventive method combined with the splitting device and the culture medium enables fast cultivation of cancer cells obtained from human tissue with a multiplication rate of 100% in all types of tumors.

Abstract: The present invention is directed to the parenteral procurement of bodily-fluid samples. The present invention is also directed to systems and methods for parenterally procuring bodily-fluid samples with reduced contamination from dermally-residing microbes. In some embodiments, a bodily-fluid withdrawing system is used to withdraw bodily fluid from a patient for incubation in culture media in one or more sample vessels. Prior to withdrawing bodily fluid into the one or more sample vessels for incubation, an initial volume of withdrawn bodily fluid is placed in one or more pre-sample reservoirs and is not used for the incubation in culture media.

Abstract: Isolated mixed populations of fetal pulmonary cells, engineered three-dimensional tissue constructs of these cells, and uses thereof in identifying therapeutic agents which augment, repair, and/or replace dysfunctional native lung and to perform in vitro studies such as pharmaceutical screening, models for lung development and disease and characterization of chemical or mechanical injury are provided.

Abstract: Methods for collecting cells in M phase or G1 phase, by which the percentage of M phase or G1 phase cells is higher than that attained by the conventional methods are disclosed.

Abstract: A process for identifying compounds that can modulate the release of neuromediators is described in which, for example, at least one compound that is to be tested is brought into contact with a nerve tissue preparation and the possible modulating effect of the compound on release of neuromediator by the nerve tissue preparation is determined. Methods of preparing calibrated pieces of mammalian cerebral material and kits for the implementation of the process are also described.

Abstract: The present invention relates to a method for inducing the differentiation of mammals' embryonic stem cells into keratinocytes, comprising the following steps of: isolating an extracellular matrix secreted by at least one mammals' cell type, cultivating mammals' embryonic stem cells in parallel in an undifferentiated condition in an appropriate culture medium in the presence of LIF, seeding the embryonic stem cells as a monolayer on said extracellular matrix, cultivating the thus seeded embryonic stem cells in the absence of LIF for a period of time sufficient for their differentiation into keratinocytes, and collecting the thus obtained keratinocytes.

Abstract: This invention relates to methods of isolating hepatoblasts utilizing panning techniques and fluorescence activated cell sorting. This invention further relates to isolated hepatoblasts and to a method of treating liver dysfunction as well as to methods of forming artificial livers.

Abstract: There is provided a method for growing human intervertebral cells. Disc tissue is surgically removed from a normal disc of a patient, the cells expanded by feeding with a cell stimulant such as a growth factor, or a cytokine or a bioactive agent to form monolayer primary cell cultures on a plastic mesh such as a nylon mesh. In the case of a growth factor, fetal bovine serum is preferred as it improves cell proliferation and production of appropriate extracellular matrix components. In another aspect of this invention, the monolayer primary cell cultures are seeded in alginate or agarose and fed again with the cell stimulant until three-dimensional cell cultures are formed. The cells are recovered from the alginate or agarose or from monolayer cultures. Re-implantation is carried out using bioresorbable carriers or cell suspensions.

Abstract: Advanced Islet Separation Technology incorporates an automated method, automated control methodology, process control interface, and automated apparatus to separate (isolate) and process pancreatic islets in a tissue suspension in physiologic process solution, utilizing microprocessor controllers and computer control and software programming to interface and control the process temperature, process flowrate, percent hydrogen concentration, dissolved oxygen concentration, endotoxin concentration, dissolved nitric oxide concentration, nitric oxide synthase concentration, proteolytic enzyme activity, and pressure of the islet containing physiologic process solution, including real-time process data acquisition and recording of the process variables.

Abstract: A method to recover receptors without removing microenvironmental components associated with them is described. The membranes containing the receptors are first associated with ligand-coupled solid supports to achieve association of the membranes through a receptor/ligand complex which includes the microenvironment to the solid support and then removing the membrane from the receptor and its microenvironment by shear forces. The thus isolated receptors and their microenvironments can then be analyzed for their role in cellular differentiation, apoptosis, transformation and the like.

Abstract: The present invention relates to a substantially pure population of viable pancreatic progenitor cells, and methods for isolating such cells. The present invention further concerns certain therapeutic uses for such progenitor cells, and their progeny.

Abstract: A device and method for capturing a biological tissue, comprises cutting a minute biological cellular tissue using a contactless cutting apparatus, wherein, by inverting a biological tissue slide provided with a sample thereon, and after cutting a cellular tissue profile using said contactless cutting apparatus, an impact lever moving mechanism applies from up to down a proper impact force or vibration force onto the target area thus cut, and thereby render said cellular tissue specimen thus captured dropping exactly through a tissue sampling hole into a sampling mortar located therebelow so as to achieve the object of capturing the desired minute biological cellular tissue.

Abstract: The present invention relates to a process for obtaining mammalian insulin secreting cells in vitro, characterized in that it contains the following steps: a) preparation of the mammalian pancreatic tissues by removal of a pancreas, b) dissociation of the pancreatic tissues obtained in step (a) into isolated pancreatic cells, c) possibly the elimination of the endocrine cells from the pancreatic cells isolated in step (b), d) induction of dedifferentiation of the cells isolated in step (b) into ductal precursor cells, e) induction of redifferentiation of the ductal precursor cells obtained in step (d) into insulin secreting cells. It also concerns the use of the insulin secreting cells thus obtained for the preparation of a pharmaceutical composition which can be used for the treatment of pancreatic pathologies and particularly diabetes.

Abstract: An apparatus and method for extracting cells from organs in which a digestion chamber includes at least one inlet and at least one outlet, and a separator for retaining the organ and permitting the cells and the physiologically compatible medium to exit the outlet. The digestion chamber can also includes at least one agitation member having an interior with at least one void. An agitation member for extracting cells from organs is also disclosed.

Abstract: A tissue harvesting method and device for obtaining micrograft tissue particles within the size range of 50-1500 microns, and more preferably 500-1000 microns, and most preferably 600 microns. The particles may be processed after a piece of donor tissue has been excised from the donor site, or processed into the desired size directly at the donor site, and thereafter excised. Cutters having blades or cutting edges spaced in the range of 50-1500 microns are utilized to obtain particles within the desired size range. An elastomer is positioned between the cutting edges to push the excised particles out of the blades for ease of use.

Abstract: The invention is directed to methods for producing a decellularized organ or part of an organ. A decellularized organ is produced using an isolated organ mechanically agitated to remove cellular membranes surrounding the isolated organ without destroying the interstitial structure of the organ. After the cellular membrane is removed, the isolated organ is exposed to a solubilizing fluid that extracts cellular material without dissolving the interstitial structure of the organ. A washing fluid is used to remove the solubilized components, leaving behind a decellularized organ.

Abstract: Demineralized bone particles are obtained by demineralizing whole bone and thereafter subdividing the demineralized bone to provide the demineralized bone particles.

Abstract: This invention relates to the establishment of a novel method of culturing sponge cells, coral cells and cells from other invertebrates in vitro. The cells cultured in vitro, which can be cultured as units similar to aggregates, are referred to as primmorphs. The method makes the following methods possible for the first time, using cells/aggregates/primmorphs from sponges, corals and other invertebrates: Methods (i) of preparing substances which modulate proliferation and DNA synthesis, (ii) of identifying/detecting environmentally harmful substances, (iii) of culturing bacteria and other micro-organisms, (iv) of preparing asexual reproductive bodies that can be used in aquaculture for growing corresponding organisms, (v) of preparing cell libraries, (vi) of optimising the nutritional requirements of the cells/aggregates/primmorphs, and (vii) of identifying substances which modulate telomerase activity in cells/aggregates/primmorphs.

Abstract: Cultures of keratinocyte cells are provided which are free from nonautologous fibroblasts and organ extracts, and which have a high speed of cell amplification for a minimum seeding density. The cultures can be cryopreserved in a buffered isotonic medium containing serum and a cryoprotectant. The cultures are produced by a process that does not involve the use of a feeder layer and organ extracts. A culture medium which can be used contains Medium 199, serum, epidermal growth factor, cholera toxin and/or hydrocortisone, and optionally insulin. A substance for wound healing and for cosmetic applications is derived from cultured human keratinocytes. A non-viable total keratinocyte lysate for use in promoting wound healing is produced by growing keratinocyte cells on a support, detaching the cells from the support, and lysing the detached cells to obtain the lysate which may be frozen and lyophilized.

Abstract: The invention is a system for maintenance and high-throughput analysis of cerebellar granule neurons in tissue culture plates under chemically defined conditions. The invention includes serum-free granule culture medium, which is composed of high glucose Dulbecco's Modified Eagle Media (DMEM), NaHCO3, sodium pyruvate, and HEPES, which is subsequently adjusted to pH 7.2. The HEPES buffered DMEM is then supplemented with L-glutamine, KCl, bovine albumin, insulin, transferrin, selenium, penicillin, and streptomycin. Unlike proprietary neuronal culture media, this invention does not include any serum, steroid hormones, phenol red, or added anti-oxidants. The serum-free granule culture medium is then placed in conventional poly-lysine coated tissue culture plates in order to conduct subsequent assays. The invention also includes the ability to package the complete neuronal culture system into a “kit” for isolation, maintenance, treatment, and analysis of cerebellar neurons.

Abstract: A method for isolating specific viable cell types from surrounding organ tissue is provided. The method entails the use of sonication in conjunction with tissue dissociating agents to free the cells of interest. A specific application of the method is the isolation of the insulin producing tissue of the pancreas, the islets of Langerhans. The method results in a high yield of islets that maintain a high level of viability.

Abstract: A method and a medium for culturing epithelial cells of both normal and malignant origin is provided. The method entails physically disaggregating tissue samples, placing the resulting fragments onto a surface comprised of basement membrane matrix components, and culturing the tissue in a medium containing preselected fetal and newborn calf sera and rat sera. Both primary explant cell cultures and cell lines, which are long-lived and particularly suitable for further study, are produced. The cultured primary explant cells undergo differentiation to form complex structures resembling those seen in vivo.

Abstract: The invention is directed to methods for producing a decellularized organ or part of an organ. A decellularized organ is produced using an isolated organ mechanically agitated to remove cellular membranes surrounding the isolated organ without destroying the interstitial structure of the organ. After the cellular membrane is removed, the isolated organ is exposed to a solubilizing fluid that extracts cellular material without dissolving the interstitial structure of the organ. A washing fluid is used to remove the solubilized components, leaving behind a decellularized organ.

Abstract: Porcine neural cells and methods for using the cells to treat neurological deficits due to neurodegeneration are described. The porcine neural cells are preferably embryonic mesencephalic, embryonic striatal cells, or embryonic cortical cells. The porcine neural cells can be modified to be suitable for transplantation into a xenogeneic subject, such as a human. For example, the porcine neural cells can be modified such that an antigen (e.g., an MHC class I antigen) on the cell surface which is capable of stimulating an immune response against the cell in a xenogeneic subject is altered (e.g., by contact with an anti-MHC class I antibody, or a fragment or derivative thereof) to inhibit rejection of the cell when introduced into the subject. In one embodiment, the porcine neural cells are obtained from a pig which is essentially free from organisms or substances which are capable of transmitting infection or disease to the recipient subject.

Abstract: This invention relates to methods of isolating hepatic progenitors utilizing panning techniques and fluorescence activated cell sorting. This invention further relates to isolated hepatic progenitors and to a method of treating liver dysfunction as well as to methods of forming artificial livers.

Abstract: The invention relates to stem cells isolated from the retina of mammals and retinal cells differentiated from these stem cells. The invention also relates to a method of isolating retinal stem cells and inducing retinal stem cells to produce retinal cells. Retinal stem cells may also be induced in vivo to produce retinal cells. The invention also includes pharmaceuticals made with retinal stem cells or retinal cells which may be used to restore vision lost due to diseases, disorders or abnormal physical states of the retina. The invention includes retinal stem cell and retinal cell culture systems for toxicological assays, for isolating genes involved in retinal differentiation or for developing tumor cell lines.

Abstract: The use of totipotent embryonic stem cells to provide substantially identical cells for embryo cloning techniques is described. The method includes the culture of loose suspensions of inner cell mass cells of bovine animals to retrieve large populations of stem cells. The invention also describes the use of stem cells in various genetic manipulation techniques.

Abstract: There is provided a method for growing human intervertebral cells. Disc tissue is surgically removed from a normal disc of a patient, the cells expanded by feeding with a cell stimulant such as a growth factor, or a cytokine or a bioactive agent to form monolayer primary cell cultures on a plastic mesh such as a nylon mesh. In the case of a growth factor, fetal bovine serum is preferred as it improves cell proliferation and production of appropriate extracellular matrix components. In another aspect of this invention, the monolayer primary cell cultures are seeded in alginate or agarose and fed again with the cell stimulant until three-dimensional cell cultures are formed. The cells are recovered from the alginate or agarose or from monolayer cultures. Re-implantation is carried out using bioresorbable carriers or cell suspensions.

Abstract: A substantially non-invasive and efficient method for collecting cells from the internal organs of a subject is provided. In one embodiment, energy from an external energy source is applied to the subject that is sufficient to loosen the cells from an internal cellular surface of an internal organ so that at least a portion of the loosened cells are detached from the internal cellular surface or the organ. The detached cells are collected from the subject and can be analyzed for a disease state. The methods described herein provide methods for detecting of disease states before macroscopic evidence of the disease state that also facilitate inexpensive mass screening.

Abstract: A perfusion device such as a liver assist device containing a housing defining a perfusion inlet and a perfusion outlet, a porous membrane structure mounted within said housing to define a perfusion compartment and an adjacent hepatocyte compartment, and porcine hepatocytes isolated from a porcine liver by retrograde perfusion.

Abstract: Novel cultured inner cell mass (CICM) cells, and cell lines, derived from ungulates, in particular, pigs and cows, and methods for their preparation are provided. The subject CICMs possess similar morphology and express cell markers identically or substantially similarly to ICMs of undifferentiated developing embryos for prolonged culturing periods. Heterologous DNA is inserted into the subject CICM cells and cell lines so as produce transgenic CICM cell which are introduced into non-human fertilized embryos to produce transgenic chimeric embryos. The transgenic chimeric embryos are transferred into recipient females where they are permitted to develop into transgenic chimeric fetuses. Recipient females give birth to transgenic chimeric animals which are capable of transmitting the heterologous DNA to their progeny. Transgenic CICM cells are also used to produce cloned transgenic embryos, fetuses and offspring.

Abstract: The present invention provides a chimeric adenovirus coat protein, which differs from the wild-type coat protein by the introduction of a nonnative amino acid sequence. Such a chimeric adenovirus coat protein according to the invention is able to direct entry into cells of a vector comprising the coat protein that is more efficient than entry into cells of a vector that is identical except for comprising a wild-type adenovirus coat protein rather than the chimeric adenovirus coat protein. The chimeric coat protein preferably is a fiber, hexon, or penton protein. The present invention also provides an adenoviral vector that comprises the chimeric adenovirus coat protein, as well as methods of constructing and using such a vector.

Abstract: A process for artificially producing three-dimensional optic tissue has been developed. The optic cells are cultured in a bioreactor at low shear conditions. The tissue forms as normal, functional tissue grows with tissue organization and extracellular matrix formation.

Abstract: Novel cultured inner cell mass (CICM) cells, and cell lines, derived from ungulates, in particular, pigs and cows, and methods for their preparation are provided. The subject CICMs possess similar morphology and express cell markers identically or substantially similarly to ICMs of undifferentiated developing embryos for prolonged culturing periods. Heterologous DNAs may be introduced into the subject CICM cells and cell lines to produce transgenic cells useful for the production of transgenic embryos, fetuses and/or offspring, e.g., by nuclear transfer procedures.

Abstract: The invention concerns a cell culture harvesting device consisting of a scraper head with a blade and a guide strip, the scraper head and the guide strip being connected with one another only by magnetic attraction. The magnetic attraction is achieved by the fact that one of the ends of the scraper head and the guide strip which are turned toward one another have a magnet and the other has either a magnet or a material which can be magnetized by the magnet of the respective counterpart. In this way, the scraper head and the guide strip can be moved synchronously and in parallel at a distance from one another. This has the advantage that the scraper head can be placed into the cell culture vessel before a cell culture is started and can be sterilized together with it thereby eliminating the risk of contamination due to a cell culture harvesting device later being placed into the cell culture vessel.

Abstract: A method for isolating specific viable cell types from surrounding organ tissue is provided. The method entails the use of sonication in conjunction with tissue dissociating agents to free the cells of interest. A specific application of the method is the isolation of the insulin producing tissue of the pancreas, the islets of Langerhans. The method results in a high yield of islets that maintain a high level of viability.

Abstract: Cultures of keratinocyte cells are provided which are free from nonautologous fibroblasts and organ extracts, and which have a high speed of cell amplification for a minimum seeding density. Preferably, a culture contains no more than about 10% autologous non-keratinocyte cells such as star-shaped, non-keratinocyte cells and no more than about 1% autologous fibroblasts. The cultures can be cryopreserved in a buffered isotonic medium containing serum and a cryoprotectant. The cultures are produced by a process that does not involve the use of a feeder layer and organ extracts. A culture medium used contains Medium 199, serum, epidermal growth factor, cholera toxin and/or hydrocortisone, and optionally insulin. A substance for wound healing and for cosmetic applications is derived from cultured human keratinocytes. Lyophilized keratinocyte cell cultures or an extract therefrom is used to provide a pharmaceutical composition. Confluent and cohesive keratinocyte sheets are prepared for use in wound healing.

Abstract: The present invention is a method for rendering cotton fiber cells that are post-anthesis and pre-harvest available for analysis of their physical properties. The method includes the steps of hydrolyzing cotton fiber cells and separating cotton fiber cells from cotton ovules thereby rendering the cells available for analysis. The analysis of the fiber cells is through any suitable means, e.g., visual inspection. Visual inspection of the cells can be accomplished by placing the cells under an instrument for detection, such as microscope or other means.

Abstract: The present invention provides a novel method for the establishment of tumorigenic cell lines from biopsies of human or animal tumors. The method includes preparing a single-cell suspension of a tumor cells by injecting cell culture medium into an isolated tumor or a portion thereof to flush out the tumor cells. The tumor or portion thereof may be in a cell culture maintenance medium. The step of injecting cell culture medium into the isolated tumor or portion thereof involves piercing the tumor or portion thereof with a needle to inject the cell culture medium and flush out the tumor cells. The steps of piercing the tumor or portion thereof with a needle and injecting the cell culture medium to flush out the tumor cells preferably are repeated until the single-cell suspension has a particular cell density. The method also includes establishing a primary tumor cell culture by growing the cells from the single-cell suspension in a cell culture medium.