35 results about "Tissue explants" patented technology

The present invention relates to new reinforced biodegradable scaffolds for soft tissue regeneration, as well as methods for support and for augmentation and regeneration of living tissue, wherein a reinforced biodegradable scaffold is used for the treatment of indications, where increased strength and stability is required besides the need for regeneration of living tissue within a patient. The present invention further relates to the use of scaffolds together with cells or tissue explants for soft tissue regeneration, such as in the treatment of a medical prolapse, such as rectal or pelvic organ prolapse, or hernia.

The present invention provides a novel method to isolate and expand pure progenitor / stem cells from a primary tissue explant, which produces a population enriched in multipotent functional progenitor / stem cells free of contaminating fibroblasts and other cell types. Cardiac progenitor / stem cells isolated by this method maintain their self-renewal and clonogenic character in vitro and differentiate into normal cells in myocardium, including cardiomyocytes, endothelial cells, and smooth muscle cells, after transplantation into ischemic hearts. The present invention also includes substantially pure populations of multipotent progenitor / stem cells, e.g., cardiac progenitor / stem cells, and their use to treat and prevent diseases and injuries, including those resulting from myocardial infarction.

The present invention provides a novel method to isolate and expand pure progenitor / stem cells from a primary tissue explant, which produces a population enriched in multipotent functional progenitor / stem cells free of contaminating fibroblasts and other cell types. Cardiac progenitor / stem cells isolated by this method maintain their self-renewal and clonogenic character in vitro and differentiate into normal cells in myocardium, including cardiomyocytes, endothelial cells, and smooth muscle cells, after transplantation into ischemic hearts. The present invention also includes substantially pure populations of multipotent progenitor / stem cells, e.g., cardiac progenitor / stem cells, and their use to treat and prevent diseases and injuries, including those resulting from myocardial infarction.

An improved method for preparing a cell culture is disclosed. The method includes culturing a multicellular tissue explant in the presence of growth medium that is substantially free of enzymes capable of digesting the explant and, subsequently, removing the explant at a predetermined time.

A method of determining at least one quantitative or qualitative pharmacological, physiological and / or therapeutic, parameter or effect of a recombinant gene product in vivo, the method comprises (a) obtaining at least one micro-organ explant from a donor subject, the micro-organ explant comprising a population of cells, the micro-organ explant maintaining a microarchitecture of an organ from which it is derived and at the same time having dimensions selected so as to allow diffusion of adequate nutrients and gases to cells in the micro-organ explant and diffusion of cellular waste out of the micro-organ explant so as to minimize cellular toxicity and concomitant death due to insufficient nutrition and accumulation of the waste in the micro-organ explant, at least some cells of the population of cells of the micro-organ explant expressing and secreting at least one recombinant gene product; (b) implanting the at least one micro-organ explant in a recipient subject; and (c) determining the at least one quantitative or qualitative pharmacological, physiological and / or therapeutic, parameter or effect of the recombinant gene product in the recipient subject.

The invention relates to a construction method and ultralow temperature freezing and storing method of a sinocyclocheilus grahami saccus olfactorius cell line, and belongs to the technical field of cell culture and ultralow temperature freezing and storing method of the biological cell of freshwater aquatic life. A sinocyclocheilus grahami saccus olfactorius tissue is used as a raw material to be cultured in an L-15 culture solution at the pH (Potential of Hydrogen) value of 7.0-7.2 and comprising fetal calf serum by means of a tissue explant and subculture is carried out by means of trypsin digestion. The method specifically comprises the preparation of a cell culture solution, primary culture and subculture. The construction method disclosed by the invention has the following beneficial effects: (1) primary culture takes short time, and cell mass is large and can be applied to chromosome analysis; (2) the constructed sinocyclocheilus grahami saccus olfactorius cell line is in fiber like morphology, can be subcultured continuously and directly applied to a research on biological characteristics, and satisfies the demands of the storage, the theoretical research and the application of sinocyclocheilus grahami germplasm resources; (3) the construction method is also applicable to the construction of saccus olfactorius cell lines of other fishes.

Reduced dyes, such as hydrocyanines, deuterocyanines, and / or other deuterated dyes capable of detecting one or more reactive oxygen species are described herein. The reduced dyes exhibit little or no fluorescence due to the disrupted π conjugation. However, upon reaction with ROS, the reduced dyes are oxidized, regenerating the extended π conjugation and causing a substantial increase in fluorescence intensity. In many case, the oxidized dye is generally membrane impermeable. However, upon reduction, many of the reduced dyes are membrane permeable. Thus, reduced dyes can accumulate in cells and / or tissue to amplify the signal. Once inside the cell or tissue, the reduced dye is reoxidized upon reaction with ROS, and the oxidized dye again becomes membrane impermeable, trapping the dye within the cell. The reduced dyes can be used to image ROS, such as hydroxide radical and superoxide, in serum, cell cultures, tissue explants, and in vivo.

The invention relates to a gastroesophageal junction smooth muscle cell tissue explant primary culture and identification method. The method includes steps: acquiring gastroesophageal junction smoothmuscle tissue muscle strips; digesting the smooth muscle tissue muscle strips by digestive juice to obtain digested smooth muscle tissue blocks; transferring the digested smooth muscle tissue blocks into a culture container to fix the digested smooth muscle tissue blocks to the bottom of the culture container, keeping a gap of 0.5-1cm between every two adjacent digested smooth muscle tissue blocks, and culturing until adherent cells occupy more than 90% of the bottom of the culture container or the adjacent digested smooth muscle tissue blocks contact through new cells. The invention further provides a smooth muscle cell identification method. The gastroesophageal junction smooth muscle cell tissue explant primary culture method is simple, perfect and effective, and specific phenotypes ofthe smooth muscle cells can be identified by adoption of the identification method.

Method for the production of taxol and / or taxanes, comprising the steps of: a) inducing the formation of callus from a plant tissue explant, through in vitro culturing in a suitable nutritient medium, b) cultivating the callus in a liquid medium to obtain a cell suspension culture capable of producing taxol and / or taxanes, c) recovering the taxol and / or the taxanes from the cells and / or from the culture medium of the cell suspension obtained from the callus in which the tissue explant is obtained from a plant of the genus Corylus, in particular Corylus avellana.

The present invention relates to methods of ex-vivo modification of mammalian tissue, via genetic transformation or introduction of cells, followed by implantation of the modified tissue into a patient in need thereof. Preferably, the tissue is microvascular free flap (or microvascular bed) tissue. A tissue explant is detached from the native circulation of a donor, transfected ex vivo, and then attached (anastomosed) to a recipient, either the donor or another patient. In a preferred embodiment, the mammalian tissue is human tissue and the patient is a human patient. Transfection with a nucleic acid encoding a product of interest is performed by contacting the selected tissue with a vector, preferably a viral vector, most preferably an adenoviral vector, that comprises the nucleic acid encoding the product of interest. The nucleic acid encoding the product of interest is driven by regulatory element such as an inducible, constitutive or cell-specific promoter, preferably an inducible or constitutive promoter. After genetic transformation of the selected tissue, the tissue is flushed to remove the vector not incorporated into the cells of the tissue. The tissue is then attached to the native circulation of the recipient using microvascular techniques. In one aspect, the invention provides methods of local delivery of a product (protein) of interest. In another aspect, the invention provides methods of systemic delivery of a product of interest. In yet another aspect, the invention provides methods of both local and systemic delivery of a product of interest. In yet another aspect, the invention provides methods for producing a "neo-organ," i.e., a non-naturally occurring vascularized tissue that provides a function of a gland or organ, or that supplements the function of a gland or organ, and that delivers locally or systemically a product of interest to a patient in need thereof.

The invention provides an establishing method of a Chinese tongue squamous cancer cell line (CTSC-1), and provides a material base for researching the characteristics of tongue squamous cancer and therapeutics thereof. The establishing method comprises the steps of: carrying out primary culture on the tongue squamous cancer cells by adopting a tissue explants adherent method; subculturing when the cell density reaches about 80 percent; carrying out RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection on stably subcultured cells to indicate that the cells express tongue squamous cancer cell symbolic molecules CK 8, 18 and 19; and detecting adherent cells through a western blot test to express proteins of the tongue squamous cancer cell symbolic molecules CK 8 and 18, therefore, proving on a molecular level the successful establishment of the CTSC-1.

The present invention provides an in vitro resveratrol-rich callus tissue of Vitis thunbergii Sieb. et Zucc. which is a callus tissue developed from a tissue culture system containing one or more plant growth regulators (PGRs) and cultured from a stem or a petiole tissue explant of a wild type of V. thunbergii or a cultivated plantlet of V. thunbergii. The cultivated plantlet is in turn derived from a shoot of the wild type of V. thunbergii cultivated in a plantlet culture system containing no PGRs. The in vitro resveratrol-rich callus tissue of V. thunbergii is characterized by its containing at least about 1,000 to 10,000 mg / kg of dried weight of resveratrol, predominantly in the form of trans-resveratrol and / or resveratrol-O-glucoside, and being ready for harvest or subculture in about 30 days. The present invention further provides a method for cultivating the in vitro resveratrol-rich callus tissue, a method for extracting the resveratrol from the in vitro resveratrol-rich callus tissue, and a method for determining the resveratrol amount in the in vitro resveratrol-rich callus tissue by HPLC.

Tissue explants of the gastrointestinal tract are provided. Methods of making and using the tissue explants are also provided, along with substrates designed for the tissue explants described.

The invention relates to construction and ultralow temperature freezing preservation method of a fin cell line of schizothorax grahami, and belongs to the technical field of culture and ultralow temperature ultralow temperature freezing preservation of cells of fresh water aquatic organisms. The method comprises the following steps of: culturing in a DMEM(Dulbecco Modified Eagle Medium) / F12 culture solution which contains fetal calf serum and cell growth factors and has the pH value of 7.0-7.2 by taking the ventral fin tissue of the schizothorax grahami as a material and adopting a tissue explant method; carrying out subculture by adopting a trypsin digestion method. The method particularly comprises the steps of cell culture solution preparation, primary culture and subculture. The construction method disclosed by the invention is short in primary culture time consumption and large in cell quantity and can be used for chromosome analysis; the constructed fin cell line of the schizothorax grahami is in a fiber-like form, can be subjected to serial passage and directly applied to biological characteristic research, meets the requirements for germplasm resource conservation and theoretical research and application of the schizothorax grahami and is suitable for constructing the fin cell lines of other fishes.

The invention discloses a method for isolating and cultivating milk cow breast cells in vitro, relating to the cell in vitro isolation and culture technology field and solving the problem that breast fibrocytes and epithelial cells of a milk cow cannot be isolated completely during the isolation and culture of the breast epithelial cells of the milk cow. The method comprises the following steps of: selecting mammary tissue of a milk cow which is killed on the very day from a local abattoir, adopting tissue explants technique and graded trypsin digestion technique to isolate and cultivate the breast epithelial cells of the milk cow according to the different sensibility of the fibroblasts and epithelial cells to trypsin and the different adhering time, purifying the breast epithelial cells of the milk cow, identifying with keratin 18 to obtain the corresponding cell system finally. The method provides the best trypsin concentration when isolating and cultivating the breast epithelial cells of a milk cow, and can completely isolate the breast fibrocytes and the epithelial cells of the milk cow from each other.

Reduced dyes, such as hydrocyanines, deuterocyanines, and / or other deuterated dyes capable of detecting one or more reactive oxygen species are described herein. The reduced dyes exhibit little or no fluorescence due to the disrupted π conjugation. However, upon reaction with ROS, the reduced dyes are oxidized, regenerating the extended π conjugation and causing a substantial increase in fluorescence intensity. In many case, the oxidized dye is generally membrane impermeable. However, upon reduction, many of the reduced dyes are membrane permeable. Thus, reduced dyes can accumulate in cells and / or tissue to amplify the signal. Once inside the cell or tissue, the reduced dye is reoxidized upon reaction with ROS, and the oxidized dye again becomes membrane impermeable, trapping the dye within the cell. The reduced dyes can be used to image ROS, such as hydroxide radical and superoxide, in serum, cell cultures, tissue explants, and in vivo.

The invention discloses a method for separating, cultivating and identifying a skin epithelial cell of a giant salamander. According to the method, a skin tissue is acquired from the tail part of the giant salamander, a tissue explant method is used to separate to acquire the primary skin epithelial cell of the giant salamander, the primary skin epithelial cell of the giant salamander is subjected to primary culture and subculture in a specific culture solution (the ingredients comprise DMEM / F12 (60%), 15% of FBS, 5 mug / mL of insulin, and 10 ng / mL of KGF) at the temperature of 28 DEG C, and specific genes (K5, K10 and P63) of the acquired skin epithelial cell of the giant salamander are expressed and identified through RT-PCR (Reverse Transcription-Polymerase Chain Reaction). The homologous design primer of the gene sequence of the related species is used to conduct PCR amplification on the genes K5, K10 and P63 of the giant salamander to acquire the gene sequences of K5, K10 and P63 of the giant salamander for the first time. The provided method for separating, cultivating and identifying the skin epithelial cell of the giant salamander lays foundations for the deep study of the skin of the giant salamander as well as the skin regeneration, trauma repair and tissue engineering skin construction with the help of the skin epithelial cell of the giant salamander.

The invention discloses a three-dimensional perfusion type cell culture device and an application thereof. The three-dimensional perfusion type cell culture device comprises a box body, a cover body and an elastic clamp, wherein a first circular groove is formed in the top surface of the box body, and a second circular groove with the same radius as the first circular groove is formed in the bottom surface of the cover body. In the cell culture device disclosed by the invention, vessels consisting of polyester fibers are arranged on the bottom surface of the groove, and a cell or tissue explant is positioned between two layers of vessels to form 'sandwich' culture; the vessels consisting of polyester fibers are used for replacing a scaffold material, so that the dead angle volume in cell culture is minimized to better conform to the in-vivo physiological microenvironment. A culture solution inlet channel and a culture solution outlet channel are formed on the side wall of the box body of the culture device respectively, and a culture solution flows into the groove cavity through the solution inlet channel and flows out through the solution outlet channel so as to realize perfusion culture. By adopting the device disclosed by the invention, the embryonic stem cell / progenitor cell at the new renal cortex part is successfully differentiated into renal tubule tissues.

The invention relates to a gene transformation method of oil sunflower, which comprises the following steps of (1) taking the tissue of a test tube plantlet of the robustly-growth oil sunflower as an explant; (2) inoculating one single bacterial colony of agrobacterium tumfaciens C58C1 into an LB liquid nutrient medium for shake cultivation overnight, taking out the bacterial colony after the OD value is 0.2-0.6 and impregnating the explant of the tissue of the test tube plantlet of the oil sunflower; (3) placing the impregnated explant of the tissue of the test tube plantlet of the oil sunflower on a differential medium containing 2-5mg / L of herbicide for differential cultivation for sub-culturing every time within 0-14 days so as to generate an inversion plantlet; (4) directly inverting the inversion plantlet screened by the herbicide into a rooting medium and transplanting the inversion plantlet into a flower pot when the inversion plantlet grows at 2-5cm. The invention establishes the set of the gene transformation method of the oil sunflower, so as to lead the steps of screening the marking concentration (the herbicide), choosing different inversion explants and adopting different sub-culturing times to be clear respectively and obtain the highest inversion efficiency, thereby laying the foundation for various gene transformations of the oil sunflower.

The invention provides an establishing method of a Chinese tongue squamous cancer cell line (CTSC-2), and provides a material base for researching the characteristics of tongue squamous cancer and therapeutics thereof. The establishing method comprises the steps of: carrying out primary culture on the tongue squamous cancer cells by adopting a tissue explants adherent method; subculturing when the cell density reaches about 80 percent; carrying out RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection on stably subcultured cells to indicate that the cells express tongue squamous cancer cell symbolic molecules CK 8, 18 and 19; and detecting adherent cells through a western blot test to express proteins of the tongue squamous cancer cell symbolic molecules CK 8 and 18, therefore, proving on a molecular level the successful establishment of the CTSC-2.

The invention provides an establishment method of a Chinese people tongue squamous epithelial cell cancer cell line and provides a material basis for research on characteristics and treatment drugs of people tongue squamous epithelial cell cancers. The establishment method comprises specific steps as follows: primary culture is performed on tongue squamous epithelial cell cancer cells with a tissue explant adherent method; subculture is performed when the cell density reaches about 80%; RT-PCR (reverse transcription-polymerase chain reaction) detection indicates that cells which are stable in subculture express people tongue squamous epithelial cell cancer cell indicative molecules CK8, 18 and 19; and western blot experiment detection indicates that adherent cells express people tongue squamous epithelial cell cancer cell indicative molecule CK8 and 18 proteins, so that the successful establishment of the Chinese people tongue squamous epithelial cell cancer cell line CTSC-1 is proved at the molecular level.

Tissue explants of the gastrointestinal tract are provided. Methods of making and using the tissue explants are also provided, along with substrates designed for the tissue explants described.

The invention discloses a method for primary culture of lotus root stem tip tissue. The method comprises the following steps: 1) collection of lotus root buds; 2) pretreatment of buds; 3) disinfection and inoculation; 4) expansion culture of shoot tip tissue; 5) leaf expansion culture. The present invention prevents browning of explant tissue by adopting suitable culture medium and culture method; prevents deformity and variation of leaves by adopting leaf-expanding culture medium, so that stems and leaves develop normally. The method can make the seedling formation rate of the top meristem of the shoot tip of the lotus root reach more than 90%.

The present invention involves a device for culturing cells on removable membranes / sponges for the purpose of transplantation. The device includes a base plate with snaps and a ring, which fits into the base plate. The membrane / sponge of choice for culturing cells is placed on the base plate and fixed by the ring, which fits tightly to the base. The device is placed inside a suitable culture vessel or receptacle and the cell suspension is added within the insert device. The cells are retained on the membrane, which enables cell attachment and proliferation. Alternatively, small tissue explants can be placed on the membrane enabling cell migration and proliferation. Cells are cultured under standard conditions with regular media changes. Following shipping to the hospitals under appropriate conditions, the insert device would be dismantled and the membrane lifted and placed at the site of transplantation.

The invention relates to a tissue block culture dish. The tissue block culture dish comprises a culture dish chassis, a culture dish top cover and a porous cover; the culture dish chassis comprises a chassis bottom surface, a chassis side surface and an 8-shaped support; the 8-shaped support comprises a support raised part, a support connection part and a support tubular part; the support tubular part is in a hollow tube structure, and the lower surface of the support tubular part is provided with the rectangular raised part; and the edge of the porous cover is provided with notches. By adopting the tissue block culture dish provided by the invention, the problem of difficult adherence to wall during tissue block culture can be solved effectively, the risk of contamination of tissue blocks can be reduced, the success rate of the tissue block culture method can be improved, and the tissue block culture can not be affected during the long-distance transfer of the tissue block culture dish.

The invention discloses a three-dimensional perfusion type cell culture device and an application thereof. The three-dimensional perfusion type cell culture device comprises a box body, a cover body and an elastic clamp, wherein a first circular groove is formed in the top surface of the box body, and a second circular groove with the same radius as the first circular groove is formed in the bottom surface of the cover body. In the cell culture device disclosed by the invention, vessels consisting of polyester fibers are arranged on the bottom surface of the groove, and a cell or tissue explant is positioned between two layers of vessels to form 'sandwich' culture; the vessels consisting of polyester fibers are used for replacing a scaffold material, so that the dead angle volume in cell culture is minimized to better conform to the in-vivo physiological microenvironment. A culture solution inlet channel and a culture solution outlet channel are formed on the side wall of the box body of the culture device respectively, and a culture solution flows into the groove cavity through the solution inlet channel and flows out through the solution outlet channel so as to realize perfusion culture. By adopting the device disclosed by the invention, the embryonic stem cell / progenitor cell at the new renal cortex part is successfully differentiated into renal tubule tissues.

The invention discloses a method for separating, cultivating and identifying a skin epithelial cell of a giant salamander. According to the method, a skin tissue is acquired from the tail part of the giant salamander, a tissue explant method is used to separate to acquire the primary skin epithelial cell of the giant salamander, the primary skin epithelial cell of the giant salamander is subjected to primary culture and subculture in a specific culture solution (the ingredients comprise DMEM / F12 (60%), 15% of FBS, 5 mug / mL of insulin, and 10 ng / mL of KGF) at the temperature of 28 DEG C, and specific genes (K5, K10 and P63) of the acquired skin epithelial cell of the giant salamander are expressed and identified through RT-PCR (Reverse Transcription-Polymerase Chain Reaction). The homologous design primer of the gene sequence of the related species is used to conduct PCR amplification on the genes K5, K10 and P63 of the giant salamander to acquire the gene sequences of K5, K10 and P63 of the giant salamander for the first time. The provided method for separating, cultivating and identifying the skin epithelial cell of the giant salamander lays foundations for the deep study of the skin of the giant salamander as well as the skin regeneration, trauma repair and tissue engineering skin construction with the help of the skin epithelial cell of the giant salamander.