Abstract: Provided is a method for efficiently and stably separating cells having a high biological activity from a biological tissue, by using a degrading-enzyme composition, which is prepared by adding an enzyme for degrading a major protein of the biological tissue in an amount determined depending on the composition of the major protein to a predetermined amount of a neutral protease and/or a protease derived from Clostridium sp. According to this method, the type and amount of protein-degrading enzyme to be used for isolating cells can be determined from the composition of a major protein of the biological tissue. Thus, cells having a high biological activity can be efficiently separated while reducing the amount of protein-degrading enzyme to be used.

Abstract: A method for the ex vivo cultivation of oral mucosal epithelial progenitor cells and oral mucosal epithelial cells includes: subjecting an oral mucosal tissue to an enzymatic digestion treatment with collagenase, so as to obtain cell aggregates which include oral mucosal epithelial progenitor cells and oral mucosal epithelial cells; cultivating the cell aggregates with an amniotic membrane in a serum-free platelet lysate-containing medium in the absence of feeder cells, so that the cell aggregates are adhered onto the amniotic membrane; and subsequently cultivating the cell aggregates adhered on the amniotic membrane in a serum-free proliferation-facilitating medium in the absence of feeder cells.

Abstract: Provided is a method for producing a glioblastoma mouse model and the mouse model produced thereby, the method including the steps of: (a) dividing a glioblastoma tissue, isolated from a patient, into 4 or more sections, and collecting one or more pieces from each of the sections; (b) dissociating a mixture of the collected pieces into glioblastoma cells as single cells; and (c) orthotopically transplanting a graft sample containing the glioblastoma cells obtained in step (b), into the brain of an immunodeficient mouse. Further provided are a method of screening a glioblastoma therapeutic agent using the mouse model and a method of providing information for selection of a patient-specific glioblastoma therapy using the mouse model. The glioblastoma mouse model shows the same genetic, morphological and pathological characteristics as those of the parental tumor, and it allows screening patient-specific glioblastoma therapeutic agent or selecting safer and more effective patient-specific glioblastoma therapy.

Abstract: Packaging for prosthetic heart valves including an assembly for stabilizing dry prosthetic tissue implants such as heart valves during storage. The packaging assembly includes a primary sterile barrier that permits gas sterilization of the tissue implant, and a secondary sterile barrier that prevents oxidation of the implant during storage. Tissue heart valves are placed inside an inner tray and sealed with a gas-permeable lid. The inner tray is placed within an outer sterile barrier and the assembly is sterilized. The outer sterile barrier may include a double seal so that a first gas-permeable seal can be closed for sterilization, after which a second gas-impermeable seal is closed to seal out any further oxygen contact with the tissue implant. Alternatively, the inner tray is placed within a sterile pouch and the assembly gas-sterilized, and then the entire assembly is placed within another pouch that provides an impermeable barrier.

Abstract: Methods of washing adherent cells, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; cell-washing solutions used for the washing method; methods of producing cell suspensions for transplantation using the cell-washing solution; and kits comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to physiological aqueous solutions to prepare cell-washing solutions containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solutions can be used to wash adherent cells before detaching the adherent cells from a culture vessel by proteolytic enzyme treatment to suppress cell death due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing the cell death due to the proteolytic enzyme treatment, such as 0.1 to 20 (w/v) %.

Abstract: Methods of washing adherent cells, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; cell-washing solutions used for the washing method; methods of producing cell suspensions for transplantation using the cell-washing solution; and kits comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to physiological aqueous solutions to prepare cell-washing solutions containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solutions can be used to wash adherent cells before detaching the adherent cells from a culture vessel by proteolytic enzyme treatment to suppress cell death due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing the cell death due to the proteolytic enzyme treatment, such as 0.1 to 20 (w/v) %.

Abstract: The invention relates to a process for generating fibroblasts, more particularly, to the culturing of fibroblasts in large numbers and of the heterogenic type. The invention is also directed to the use of fibroblasts in the preparation of heterotypic spheroids and a process for the preparation of such heterotypic spheroids.

Abstract: A process of diagnostic, prognostic and therapeutic monitoring of solid tumors, and new biological markers of tumor.

Abstract: In a process for preparing disinfected preparations from mammalian tissues, mammalian tissue is perfused at one step with an enzyme-free liquid antibiotic composition that includes at least one antibiotic and is contained in at least one perfusion buffer. In a step occurring after such one step, enzymatic antibiotic-free treatment of the tissue is carried out to obtain a single-cell suspension. In another step disinfected preparations are obtained.

Abstract: In a process for preparing disinfected preparations from mammalian tissues, mammalian tissue is perfused at one step with an enzyme-free liquid antibiotic composition that includes at least one antibiotic and is contained in at least one perfusion buffer. In a step occurring after such one step, enzymatic antibiotic-free treatment of the tissue is carried out to obtain a single-cell suspension. In another step disinfected preparations are obtained.

Abstract: The invention provides a method of isolating dermal stem cells, having the steps of subjecting cells separated from the skin by enzyme treatment to suspension culture, and isolating cells positive for stem cell markers from the cultured cells.

Abstract: A method for suppressing an immune response is provided. The method involves administration of isolated lymphoid tissue-derived suppressive stromal cells (LSSC) to a subject in need of such treatment in an amount effective to suppress the immune response in the subject. The invention also involves a method to isolate LSSC by digesting lymphoid tissue fragments using a combination of an enzyme mix and agitation and then collecting the LSSC. Pharmaceutical preparations comprising LSSC are also provided.

Abstract: A method of preparing a stem cell preparation is provided. The method includes steps of collecting adipose tissue from a mammal, contacting that adipose tissue with an enzyme preparation to digest fat and connective tissue and preserve stem cells and collecting those stem cells. In addition a kit is provided for preparing a stem cell preparation.

Abstract: Methods of manufacturing a bone-derived extracellular matrix (bECM) product are provided. Also provided are bECM products and methods of using those products.

Abstract: An isolated mammalian internal mammary artery-derived cell is disclosed. Furthermore, methods of isolating the mammalian internal mammary artery-derived cell are disclosed. The cell is useful in tissue engineering technologies, specifically in vascular tissue engineering.

Abstract: Provided herein are devices and methods for the micro-isolation of biological cellular material. A micro-isolation apparatus described can comprise a photomask that protects regions of interest against DNA-destroying illumination. The micro-isolation apparatus can further comprise photosensitive material defining access wells following illumination and subsequent developing of the photosensitive material. The micro-isolation apparatus can further comprise a chambered microfluidic device comprising channels providing access to wells defined in photosensitive material. The micro-isolation apparatus can comprise a chambered microfluidic device without access wells defined in photosensitive material where valves control the flow of gases or liquids through the channels of the microfluidic device.

Abstract: Adipose tissue-derived stromal cells and methods of isolating and using the same. In at least one embodiment of isolated adipose tissue-derived stromal cells of the present disclosure, the cells are isolated by performing adipose tissue resection or suction on a mammalian patient, dissecting tissue obtained from said tissue resection or suction and dissociating said tissue into a cell suspension, removing adipocytes from the cell suspension, culturing the adipocyte-depleted cell suspension in EGM-2-MV media, and isolating adipose tissue-derived stromal cells secreting vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and granulocyte-colony stimulating factor (G-CSF).

Abstract: Disclosed are perivascular medicinal cells (PVMCs), methods of isolating PVMCs, and a composition comprising PVMCs having medicinal capabilities. Specifically, the disclosure provides a method of isolating PVMCs from an umbilical cord blood vessel or from bone. The disclosure further provides a method of making an enhanced, autologous bone graft, a method of stimulating bone regeneration comprising administering a therapeutically effective amount of bone-derived PVMCs, a method of reconstructing bone tissue, and methods of treating a disease that affects cellular function. PVMCs capable of secreting a site-dependent trophic factor are also described.

Abstract: A method of obtaining stromal progenitor cells (SPC) from subcutaneous adipose tissue by incubation of a very small volume of the subcutaneous adipose tissue in an enzyme solution produces SPC that are usable in medical applications based on autologous SPC even on individuals having a body mass index lower than 18.5.

Abstract: Method for the isolation, expansion and preservation of cardiac stem cells from human or animal tissue biopsy samples to be employed in cell transplantation and functional repair of the myocardium or other organs. Cells may also be used in gene therapy for treating cardiomyopathies, for treating ischemic heart diseases and for setting in vitro models to study drugs.

Abstract: Disclosed is a novel cell mass derived from a cancer tissue, which can reflect the in vivo behavior of a cancer cell correctly. Also disclosed is a process for preparing the cell mass. Specifically disclosed is a cell mass derived from a cancer tissue, which is an separated product that is isolated from a cancer tissue obtained from an individual as a mass containing at least three cancer cells or a cultured product of the separated product and which can retain a proliferation ability in vitro. The cell mass derived from a cancer tissue is produced by, for example, a preparation process comprising the steps of: treating a pulverized product of a cancer tissue removed from a living body with an enzyme; and selecting and collecting a mass containing at least three cancer cells among from an enzymatic treatment product.

Abstract: A unitary apparatus for isolating cells from adipose tissue including a lipid separation processor with a dispersing head equipped with a plurality of ports and a digestion chamber for dissociation of the constituent cells disposed in adipose tissue. The lipid separating apparatus is useful for the separation of lipids and adipocytes from a mixed cell population. A cell seeding chamber may be attached to the cell isolation apparatus. The components of the apparatus may be packaged in modular kit form.

Abstract: The invention is directed to methods and compositions for obtaining uniform sized muscle fiber fragments for transplantation. These muscle fiber fragments are able to reconstitute into long fibers that are oriented along native muscle. The implanted muscle cells integrate with native vascular and neural network, as confirmed by histology and immunohistochemistry. This invention is particularly advantageous because autologous muscle can be harvested from a donor site, processed and injected into target sites in the operating room. The fragmented muscle fibers can be readily integrated within the host.

Abstract: Disclosed is a novel cell mass derived from a cancer tissue, which can reflect the in vivo behavior of a cancer cell correctly. Also disclosed is a process for preparing the cell mass. Specifically disclosed is a cell mass derived from a cancer tissue, which is an separated product that is isolated from a cancer tissue obtained from an individual as a mass containing at least three cancer cells or a cultured product of the separated product and which can retain a proliferation ability in vitro. The cell mass derived from a cancer tissue is produced by, for example, a preparation process comprising the steps of: treating a pulverized product of a cancer tissue removed from a living body with an enzyme; and selecting and collecting a mass containing at least three cancer cells among from an enzymatic treatment product.

Abstract: Cells derived from postpartum umbilicus and placenta are disclosed. Pharmaceutical compositions, devices and methods for the regeneration or repair of ocular tissue using the postpartum-derived cells are also disclosed.

Abstract: Method for the isolation, expansion and preservation of cardiac stem cells from human or animal tissue biopsy samples to be employed in cell transplantation and functional repair of the myocardium or other organs. Cells may also be used in gene therapy for treating cardiomyopathies, for treating ischemic heart diseases and for setting in vitro models to study drugs.

Abstract: Disclosed herein are compositions and methods for treating, ameliorating or preventing a retinal disease or condition; improving a photopic (day light) vision; for improving correcting visual acuity, improving macular function, improving a visual field, or improving scotopic (night) vision by administration of retinal progenitor cells. The subject matter described herein also provides cell populations comprising retinal progenitor cells and methods of isolation thereof.

Abstract: The use of a neutral protease (NP) together with a collagenase consists in that a neutral protease which is not contained in a collagenase enzyme preparation and which is not produced by a recombinant production is mixed before the beginning of a tissue dissociation with a collagenase or a collagenase enzyme preparation with an individual dosage of the quantitative proportions of neutral protease and collagenase for improving the isolation results with respect to yield, viability and integrity of the cells.

Abstract: The disclosed islet isolation method comprises: an injection step of injecting a preservation solution into the pancreatic duct of an excised pancreas; a preservation step of immersing the pancreas into an immersion fluid for preservation; a digestion step of breaking down the pancreas to provide pancreatic tissue; and a purification step of immersing the pancreatic tissue in a purification solution to provide islets. The digestion step consists of: an enzyme injection step of injecting an enzyme solution containing a digestion enzyme into the pancreas; a digestion initiation step of activating the digestion enzyme; a digestion termination step of inactivating the digestion enzyme; and a collection step of collecting the broken-down pancreatic tissue.

Abstract: Disclosed herein are compositions and methods useful for preparing neural scaffolds. The scaffolds comprise tissue taken from the spinal cord and/or dura mater of vertebrate and can be processed to form gels or sheets. Methods of treating patient with CNS injury are also presented.

Abstract: Provided herein is a placental product comprising an immunocompatible chorionic membrane. Such placental products can be cryopreserved and contain viable therapeutic cells after thawing. The placental product of the present invention is useful in treating a patient with a tissue injury (e.g. wound or burn) by applying the placental product to the injury. Similar application is useful with ligament and tendon repair and for engraftment procedures such as bone engraftment.

Abstract: The present invention relates to testis somatic cell-derived pluripotent stem cells, and more particularly, to pluripotent adult stem cells which exhibit a positive immune reaction to both CD34 and CD73 and which are derived from testis somatic cells. The present invention further relates to a method for producing the testis somatic cell-derived pluripotent stem cells, and to a pharmaceutical composition including same for the treatment of erectile dysfunction.

Abstract: Cells derived from postpartum tissue and methods for their use in treatment of diseases of the heart or circulatory system are disclosed. Cells may be used in either differentiated or undifferentiated forms, in homogenous cultures, or as populations with other cells, and in conjunction with other bioactive factors.

Abstract: The invention relates to a method for isolating cells from a tissue sample. In preferred embodiments, chondrocytes are isolated from cartilage tissue in a shorter time than hitherto considered possible.

Abstract: This invention is directed to methods of directly reseeding harvested adherent cells grown in a hollow fiber bioreactor. Also disclosed is a novel harvest media for use in directly reseeding adherent cells into a hollow fiber bioreactor.

Abstract: Methods of washing adherent cells, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; cell-washing solutions used for the washing method; methods of producing cell suspensions for transplantation using the cell-washing solution; and kits comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to physiological aqueous solutions to prepare cell-washing solutions containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solutions can be used to wash adherent cells before detaching the adherent cells from a culture vessel by proteolytic enzyme treatment to suppress cell death due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing the cell death due to the proteolytic enzyme treatment, such as 0.1 to 20 (w/v)%.

Abstract: Disclosed is a method for isolating stem cells. The method uses a specially designed apparatus including: a container containing an aspirate; a piston having an outer diameter corresponding to the inner diameter of the container and having at least one through-hole; and a connection tube adapted to feed an enzyme or a washing solution into the container through the through-hole, having a tip connected to the through-hole, and connected to an external tube or another container containing the enzyme or washing solution at the other end thereof. The method includes pulling the piston backward to form a negative pressure in the container containing the aspirate and to allow the enzyme or washing solution to enter the container containing the aspirate through the connection tube and the through-hole of the piston.

Abstract: Provided herein are novel methods and compositions utilizing adipose tissue-derived stromal stem cells for treating fistulae.

Abstract: Populations enriched for smooth muscle progenitors are obtained by selection on the basis of expression of specific cell surface markers.

Abstract: There is disclosed a method of combining mesenchymal stem cells (MSCs) with an osteochondral allograft. In an embodiment, the method includes obtaining adipose tissue having MSCs with unwanted cells. The tissue is digested to form a cell suspension having MSCs and unwanted cells. The suspension is added to seed the allograft. The MSCs are allowed to attach to the allograft. There is disclosed an allograft product including MSCs with an osteochondral allograft. There is disclosed a method of combining MSCs with decellularized, morselized cartilage. In an embodiment, the method includes obtaining adipose tissue having MSCs with unwanted cells. The tissue is digested to form a cell suspension having MSCs and unwanted cells. The suspension is added to seed the cartilage. The MSCs are allowed to attach to the cartilage to. There is disclosed an allograft product including MSCs with morselized cartilage. Other embodiments are also disclosed.

Abstract: The invention provides a method of isolating dermal stem cells, having the steps of subjecting cells separated from the skin by enzyme treatment to suspension culture, and isolating cells positive for stem cell markers from the cultured cells.

Abstract: The present invention relates to a method for isolating stem/progenitor cells from the amniotic membrane of umbilical cord, wherein the method comprises separating the amniotic membrane from the other components of the umbilical cord in vitro, culturing the amniotic membrane tissue under conditions allowing cell proliferation, and isolating the stem/progenitor cells from the tissue cultures. The isolated stem cell cells can have embryonic stem cell-like properties and can be used for various therapeutic purposes. In one embodiment, the invention relates to the isolation and cultivation of stem cells such as epithelial and/or mesenchymal stem/progenitor cells under conditions allowing the cells to undergo mitotic expansion. Furthermore, the invention is directed to a method for the differentiation of the isolated stem/progenitor cells into epithelial and/or mesenchymal cells.

Abstract: This invention relates to a method for the reconstruction of a tissue-engineered human corneal endothelium. Human corneal endothelial cells are cultured in vitro to logarithmic growth phase using 20% calf bovine serum-containing DMEM/F12 medium. Trypsin is used to digest epithelial layer of the freeze-dried human amniotic membrane in order to produce denuded human amniotic membrane as scaffold carriers. The scaffold carriers are tiled on the bottom of culture plate wells until they are dried and completely adhered to the bottom of wells. Human corneal endothelial cells at logarithmic growth phase are re-suspended in DMEM/F12 medium containing type-IV collagen and 20% calf bovine serum. Human corneal endothelial cell suspension is subsequently inoculated to amniotic membrane scaffold carriers that are tiled on the bottom of wells in culture plate to launch in vitro culture as well as in vitro reconstruction of tissue-engineered human corneal endothelium. This invention is scientific and rational.

Abstract: The invention provides methods for diagnosing diseases such as cancer and other conditions using biological samples. Liquid Tissue samples prepared from histopathologically prepared tissue obtained from a subject surprisingly can be used to identify and, optionally, to quantify analytes that are diagnostic of the presence of a disease, condition or syndrome in the subject.

Abstract: Solutions and suspensions comprising polymerized hemoglobin derived from human blood are disclosed. The solutions and suspensions may comprise cell culture medium, an enzyme (such as a protease), and/or a buffer. Processes of preparing the solutions and suspensions are also disclosed. The solutions and suspensions may be employed in methods of isolating mammalian cells, such as pancreatic islets, methods of preserving mammalian tissue and organs, methods of aiding the recovery of mammalian cells following their isolation, methods of maintaining mammalian cells, methods of propagating mammalian cells, and methods of treating a mammal with diabetes.

Abstract: Compositions useful for treatment of patients needing hematopoietic stimulation. In one embodiment patients are administered a cellular mixture derived from allogeneic placenta, said cellular mixture comprising substantially of endothelial cells and endothelial progenitor cells.

Abstract: The present invention is a process for preparing soft tissue such as tendons, ligaments, cartilage, fascia, dermis, human valves and human veins for implant in a human and removes cellular components and forms an decellular matrix having as major components collagens and elastins while sterilizing the tissue. The process comprises the following steps: (1) isolating from a suitable donor a desired soft tissue sample of the biological material; (2) processing and decellularizing the soft tissue including inspection for visual defects, trimming and soaking the tissue in a detergent depending on whether the tissue is fascia or dermis and rinsing same with sterile water; (3) sterilizing the soft tissue in a vacuum and soaking the tissue in an antibiotic composition or peracetic acid depending on whether the soft tissue is fascia or dermis and rinsing same; (4) processing the tissue by cutting the tissue to size and perforating the tissue; and (5) dipping the tissue in 70% ethanol and packaging the tissue.

Abstract: The present invention contemplates the creation of a low fluoride oil processed from a phospholipid-protein complex (PPC) formed immediately upon a crustacean (i.e., for example, krill) catch. The process comprises disintegrating the crustaceans into smaller particles, adding water, heating the result, adding enzyme(s) to hydrolyze the disintegrated material, deactivating the enzyme(s), removing solids from the enzymatically processed material to reduce fluoride content of the material, separating and drying the PPC material. Then, using extraction with supercritical CO2 and ethanol as solvents, inter alia krill oil is separated from the PPC. In the extraction the krill oil can be separated almost wholly from the feed material. The products have low fluoride content. The manufacturing costs in the extraction process are relatively low.

Abstract: The invention provides enzymatic methods for direct determination of percentage of glycated hemoglobin in blood samples without the need of a separated measurement of total hemoglobin content in blood samples. The methods utilizes one or two different types of oxidizing agents which selectively oxidize low-molecular weight reducing substances and high-molecular weight (mainly hemoglobin) reducing substances in blood samples, coupled with enzymatic reactions catalyzed by proteases, fructosyl amino acid oxidase. The amount of hydrogen peroxide generated in the reaction is measured for determination of percentage of glycated hemoglobin in blood samples. The invention provides kits for performing the methods of the invention.

Abstract: Those provided here are an Activin A-containing solution for making inhibitory neuron progenitors proliferate, a method for culturing the inhibitory neuron progenitors using this solution, a method for separating the inhibitory neuron progenitors and the inhibitory neurons using an Activin A receptor Acvr1 expression as an index, a method for transplanting thus separated cells to the brain surface, and a method for screening a factor for making the inhibitory neuron progenitors proliferate.