139 results about "Feeder Layer" patented technology

Methods and materials for culturing primate-derived primordial stem cells are described. In one embodiment, a cell culture medium for growing primate-derived primordial stem cells in a substantially undifferentiated state is provided which includes a low osmotic pressure, low endotoxin basic medium that is effective to support the growth of primate-derived primordial stem cells. The basic medium is combined with a nutrient serum effective to support the growth of primate-derived primordial stem cells and a substrate selected from the group consisting of feeder cells and an extracellular matrix component derived from feeder cells. The medium further includes non-essential amino acids, an anti-oxidant, and a first growth factor selected from the group consisting of nucleosides and a pyruvate salt.

The invention relates to methods for culturing human embryonic stem cells by culturing the stem cells in an environment essentially free of mammalian fetal serum and in a stem cell culture medium including amino acids, vitamins, salts, minerals, transferring, insulin, albumin, and a fibroblast growth factor that is supplied from a source other than just a feeder layer the medium. Also disclosed are compositions capable of supporting the culture and proliferation of human embryonic stem cells without the need for feeder cells or for exposure of the medium to feeder cells.

The invention relates to methods for culturing human embryonic stem cells by culturing the stem cells in an environment essentially free of mammalian fetal serum and in a stem cell culture medium including amino acids, vitamins, salts, minerals, transferring, insulin, albumin, and a fibroblast growth factor that is supplied from a source other than just a feeder layer the medium. Also disclosed are compositions capable of supporting the culture and proliferation of human embryonic stem cells without the need for feeder cells or for exposure of the medium to feeder cells.

The present invention provides a non-mouse, including human, pluripotential embryonic stem cell which can:(a) be maintained on feeder layers for at least 20 passages; and(b) give rise to embryoid bodies and multiple differentiated cell phenotypes in monolayer culture.The invention further provides a method of making a pluripotential embryonic stem cell comprising culturing germ cells and germ cell progenitors in a composition comprising a growth enhancing amount of basic fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor to primordial germ cells under cell growth conditions, thereby making a pluripotential embryonic stem cell.Also provided are compositions useful to produce the pluripotent embryonic stem cells and methods of screening associated with the method of making the embryonic stem cell.

The present invention provides compositions and methods for the production of differentiated mammalian cells. More particularly, the present invention provides cellular differentiation methods employing culturing the cells on a feeder layer or under feeder-free conditions in cell culture and further contacting the cells with an inhibitor of the PI3-kinase pathway for the generation of differentiated mammalian cells from pluripotent mammalian stem cells. Preferably, the differentiated cell is selected from the group consisting of a mesendodermal cell, a mesodermal cell, and an endodermal cell.

Methods and composition of induction of pluripotent stem cells are disclosed. For example, in certain aspects methods for generating essentially vector-free induced pluripotent stem cells with cell signaling regulators are described. Furthermore, certain aspects of the invention provide novel compositions comprising induced pluripotent stem cells essentially free of exogenous retroviral vector elements in the presence of a medium comprising signaling inhibitors. In certain aspects, feeder-free episomal reprogramming methods may be provided.

The current invention relates to the control and / or manipulation of the gamma-secretase signaling pathway in pluripotent cells to stabilize the cells in a pluripotent state and / or to control the differentiation of the pluripotent cells towards a differentiated state. The invention further includes feeder layers that contain or express ligands or other compounds that inhibit gamma-secretase or Notch signaling to enhance the maintenance of pluripotent cells in a pluripotent state. The invention also includes cell culture compositions that comprise pluripotent cells and inhibitors of gamma-secretase, or activators or inhibitors of Notch signaling.

Described are methods for isolating / purifying and expanding animal stem cells and stem-cell-like cells. Isolation methods include conditions comprising preferentially digesting non-stem cells and non-stem-cell-like cells in a population and preferentially adhering stem cells and stem-cell-like cells in a population. Expansion methods include culturing such cells under conditions comprising modulation of TGF-β signaling, inhibition of cell signaling mediated by p38 MAP kinase using small molecular weight inhibitors, expansion of the cells on human amniotic epithelial cells as feeder layers, control of cell seeding density, control of levels of Ca2+ in the culture media, rapid adhesion on a substrate or by a combination of such conditions. More particularly, what is disclosed relates to methods and systems for expanding animal cells in ex vivo cell cultures, while preventing cellular differentiation, and selectively enriching stem cells. The embodiments also disclose a culture system for ex vivo expansion of limbal epithelial cells or mesenchymal cells, as well as surgical grafts made there from.

A dynamic layer diagnostic device for smart power grid fault comprises a data collection and monitoring unit, a data processing unit, a data base unit, a communication unit and a man-machine interaction unit. A dynamic layer diagnostic method includes: when a smart power grid breaks down, calculating fault diagnosis starting conditions to conform a diagnosis strategy, wherein the fault diagnosis strategy comprises switch layer diagnosis, feeder layer diagnosis and transformer substation layer diagnosis; starting the switch layer diagnosis when changes of switch motion information are remarkable before and after the fault; starting the transformer substation diagnosis when changes of electricity amount information are remarkable before and after the fault; stopping the diagnosis when a fault element is the only one element during the diagnosis of the switch layer; otherwise, retrieving the switch historical action recording, and starting the transformer substation diagnosis when recording matched with the current switch action exists; and otherwise, starting the feeder layer diagnosis. The dynamic layer diagnostic method performs layering analysis on the fault, fully utilizes various fault information and improves fault diagnosis accuracy according to different characteristics of multisource information after the power grid fault and difficulty layer in obtaining and processing of various information.

The present invention provides compositions and methods for the culture and maintenance of pluripotent stem cells. More particularly, the present invention provides for compositions and methods for culturing, maintaining, growing and stabilizing primate pluripotent stem cells in a feeder-free defined media further comprising human serum, or a soluble attachment component of the human serum, for promoting cell attachment.

The present invention relates to methods for producing feeder cell-free neuroprogenitor cells (preferably adherent) from embryonic stems cells, preferably human embryonic stem cells, the feeder cell-free neuroprogenitor cells, preferably human cells themselves, as well as methods for producing feeder cell-free samples of neuronal cells, preferably adherent human neuronal cells and the feeder cell-free neuronal cells themselves. Pharmaceutical compositions and methods of treating neurodegenerative diseases as well as the use of the described cells in assay systems is also described.

The invention relates to methods for culturing human embryonic stem cells by culturing the stem cells in an environment essentially free of mammalian fetal serum and in a stem cell culture medium including amino acids, vitamins, salts, minerals, transferring, insulin, albumin, and a fibroblast growth factor that is supplied from a source other than just a feeder layer the medium. Also disclosed are compositions capable of supporting the culture and proliferation of human embryonic stem cells without the need for feeder cells or for exposure of the medium to feeder cells.

A hierarchy of endothelial colony forming cells (EPCs) was identified from mammalian cord blood, umbilical vein and aorta. A newly isolated cell named high proliferative potential—endothelial colony forming cell (HPP-ECFC) was isolated and characterized. Single cell assays were developed that test the proliferative and clonogenic potential of endothelial cells derived from cord blood, or from HUVECs and HAECs. EPCs were found to reside in vessel walls. Use of a feeder layer of cells derived from high proliferative potential-endothelial colony forming cells (HPP-ECPCS) from human umbilical cord blood, stimulates growth and survival of repopulating hematopoietic stem and progenitor cells. Stimulation of growth and survival was determined by increased numbers of progenitor cells in in vitro cultures and increased levels of human cell engraftment in the NOD / SCID immunodeficient mouse transplant system.

Improvements are made to a novel media that replace the requirement for all protein growth factors by the addition to the medium of physiological concentrations of retinyl acetate. The media are serum-free, companion cell or feeder layer-free and organotypic, matrix free solutions for the cultivation of clonally competent basal keratinocytes. The media and methods are useful in the production of epidermal epithelial tissue that is suitable for skin grafting.

The invention discloses a high-density integration tile-type active phased-array antenna structure, and aims to provide a high-integration tile-type active phased-array antenna structure which is smaller in volume, lighter in weight, lower in cost and stable in performance and can eliminate a self-excitation phenomena. The structure is realized through the following technical scheme: a radiation array element layer is arranged on an upper multi-layer PCB (2), each metal radiation patch (1) array module is electrically connected to an MMIC chip (5) through a radio-frequency feeder line (4), the MMIC chips are packaged in metal sealed cavities (12) between the upper multi-layer PCB and a lower multi-layer PCB to form the RF feeder layer; each MMIC chip is electrically connected to a channel assembly (7), a power supply assembly (8) and a beam control assembly (9) which are positioned on the lower bottom plane of a multi-layer printed board (3) respectively, and an DC power supply and a control circuit layer are formed by the MMIC chips as well as a radio-frequency external interface (10) and a control and power supply external interface (11) which are arranged at two ends of the bottom of the multi-layer printed board respectively.

The invention provides cell culture conditions for culturing stem cells, including feeder-free conditions for generating and culturing human induced pluripotent stem cells (iPSCs). More particularly, the invention provides a culture platform that allows long-term culture of pluripotent cells in a feeder-free environment; reprogramming of cells in a feeder-free environment; single-cell dissociation of pluripotent cells; cell sorting of pluripotent cells; maintenance of an undifferentiated status; improved efficiency of reprogramming; and generation of a naïve pluripotent cell.

The invention discloses a low-sidelobe antenna array based on a micro-strip ridge gap waveguide unequal power divider network. The low-sidelobe antenna array is composed of a radiation layer dielectric plate, a gap layer, and a feeder layer dielectric plate. An electrical wall formed by plated through holes in a radiation dielectric layer and 2*2 radiation slots constitute back cavity antenna sub-arrays, and the sub-arrays are arranged periodically to form a radiation antenna array. An underlying unequal power divider network feeds power to the radiation antenna array via a coupling slot. There is an air gap of certain height between the feeder layer dielectric plate and the radiation layer dielectric plate. By using the low-sidelobe antenna array based on a micro-strip ridge gap waveguide unequal power divider network, the amplitude weighted distribution and phase matching problems of the micro-strip ridge gap waveguide unequal power divider network are solved. Moreover, the low-sidelobe antenna array has such advantages as simple structure, easy integration, simple processing, small size, wide bandwidth, low insertion loss and stable structural performance.

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. The cells may be grown without using a support to produce the lysate, or to produce a non-viable keratinocyte cell culture lyophilisate or spray dried non-viable keratinocyte cell composition for use in healing wounds.

A method of producing a semiconductor device having a heat dissipating metal layer wherein the number of patterning steps is reduced, laser dicing produces a better profile, and first and second metal layers are prevented from separating from each other, and a semiconductor device produced by the method. The number of patterning steps is reduced by employing a flat exposure step for photoresist with mask alignment. A better appearance is obtained by forming the metal layers which connect the semiconductor devices with each other from a first metal layer having a lower melting point and a second metal layer having a higher melting point and severing the first metal layer and the second metal layer successively, from the first metal layer side. A second metal layer is prevented from peeling by preventing oxidation of the plated feeder layer through plating of the second metal layer.

The invention discloses a six-tower distilling device and technology of superfine edible spirit, which comprises the following parts: crude distillation column, vacuum aldehydo-removing column, water scrubber, fractionating tower, methanol column and recycling tower, wherein the mature ferment fermented glutinour rice is preheat in the ferment procedure, which enters into the top of crude distillation column; the crude distillation column is negative pressure, whose top wine gas enters in the vacuum aldehydo-removing column through separator after cooling sequently; the aldehydo-removing wine on the bottom is sent into feeder layer of water scrubber after preheating, which flows desalination wine after extracted by micro-negative pressure water in the fractionating tower to proceed positive pressure distillation; the spirit intermediate product is led into the methanol column from top wine hole of fractionating tower to remove menthol, which obtains superfine edible spirit; the steam condensate of fractionating tower reboiler is recycled in the autoclave of recycling tower after flash-boiling disposal, which flows through methanol column reboiler, water scrubber and crude distillation column to proceed secondary condensing by reboiler.

The invention relates to a multilayer micro-strip flat-plate array antenna with symmetric beams, which consists of a radiation layer, a feeder layer and a reflection plate, wherein radiation patches are symmetrically distributed on an inner surface of the radiation layer, and the radiation patch array is totaled in 18 units of 3*6; the feeder layer is arranged between the radiation layer and the reflection plate, a distance between every two adjacent layers is a quarter wavelength, a grounding surface of the feeder layer is provided with 18 H-shaped coupled gaps in a form of 3*6, a micro-strip feeder line is arranged on the feeder surface of the feeder layer, on a micro-strip feeder-line network, energy is allocated by a first-level power distributor to a second-level power distributor and is opposite to a phase position, finally the energy is allocated to each radiation unit through a serial impedance matching circuit, the reflection plate is a metal plate and forms an inner surface of an antenna shell, the radiation layer also has a function of an antenna cover, and the entire antenna is completely arranged inside a closed shell except a connector.

The invention provides an automatic chicken-feet cutting equipment complex technical proposal capable of safely, effectively and automatically cutting the chicken-feet, characterized in that: the equipment complex comprises a vibration feeder, a material conveying device and a pressure device; the vibration feeder layers the chicken-feet on the conveyer belt of the material conveying device; the material conveying device cooperates with the pressure device and the conveyer belt intermittently moves to convey the chicken-feet on a mould cutting area of the pressure device. Thus the equipment complex is set in continuous punching mode when starting the pressure device, thereby safely, effectively and automatically cutting the chicken-feet.

The present invention provides a culture substrate which enables maintenance culture of human pluripotent stem cells in a pluripotent state under a feeder-free culture environment, and a culture method of human pluripotent stem cells using the culture substrate. By seeding human pluripotent stem cells dissociated into single cells at a cell density of 4×104 to 10×104 cells / cm2 onto a culture substrate coated with human laminin α5β1γ1 E8 fragment or human laminin α3β3γ2 E8 fragment preferably at a concentration of 0.5 to 25 μg / cm2, the human pluripotent stem cells can be rapidly expanded in a pluripotent state.

The invention discloses an optical fiber composite power seabed oil-filled cable, which is characterized in that a light unit layer is arranged in a defending layer. The light unit layer is twisted by an optical fiber and polyethylene filled bar at interval. A metal protecting layer comprises a lead protecting layer, wherein a return feeder layer is arranged outside the lead protecting layer. The return feeder layer is assembled by a flat copper line. A lining layer is arranged between the lead protecting layer and the return feeder layer. The return feeder layer is arranged in the metal protecting layer so as to improve the short circuit current of the metal protecting layer and help to improve the current carrying amount of the cable at the same time. The invention has the advantages of higher electrical performance and sufficient mechanical intensity by combining the seabed optical fiber cable and seabed power cable.

The present invention provides compositions and methods for the culture and maintenance of cancer stem cells. More particularly, the present invention provides the identification of cancer stem cell specific markers and methods of recognizing the same for the detection of tumors, for facilitating the prognosis of a patient with a tumor, and for the treatment of various cancers. The invention also provides antibodies that specifically recognize the disulfide linked Erbb2Δ16 homodimer, an Erbb2Δ16 / Erbb3 heterodimer, or post-translational modifications of Erbb2 that are specific to Erbb2 of variant hESCs. In addition, the invention provides a modified defined media useful in the absence of a feeder layer and in the absence of serum or serum replacement, that comprises a basal salt nutrient solution, bFGF, IGF-I, and Activin A, and wherein the composition does not comprise heregulin. The invention further relates to the use of an Erbb2 variant isoform to generate robust cell cultures that are independent of heregulin.

The invention discloses a method and a special culture medium for subculturing chicken embryonic stem cells for a long time. The method is characterized by comprising the following steps of: isolating cells of area pellucida of X-stage blastoderm, dispersing into single cells or small cell masses through mechanical blowing and beating, inoculating on an STO feeder layer, culturing the chicken embryonic stem cells in the special culture medium at the temperature of between 37 and 38 DEG C, and subculturing once every 3 to 5 days, wherein the special culture medium comprises 600 to 900mL of conditioned medium, 50 to 150mL of fetal calf serum, 5 to 20mL of chicken serum, 5 to 25mL of one or a mixture of more of non-essential amino acids, 0.146 to 0.292g of L-glutamine, 6 to 14mu L of beta-mercaptoethanol, 1 to 2*10<6> IU of mouse leukemia inhibitory factor, 10 to 50mu g of alkaline fibroblastic growth factor and 5 to 20mu g of stem cell growth factor; and fixing the volume of the ingredients to 1,000mL by using a dulbecco's modified eagle medium (DMEM) (high glucose), and regulating the pH value to 7.2 to 7.5. The conditioned medium is obtained by the steps of: culturing BRL-3A cells until the cells are converged, collecting supernatant of the cultured cells, performing centrifugal separation, and filtering by using a filter membrane to obtain the conditioned medium. Compared with the prior art, the method has the advantages that: the long-term subculture of the chicken embryonic stem cells can be realized, the proliferation of the cells is quick, and the cloning yield is high.

The invention provides a culture composition for amplifying primary NK cells in vitro without a feeder layer and the application thereof, and relates to obtained NK cells and related application. Theinvention provides a composition for culturing the NK cells. The composition comprises an activating culture medium composition and / or an amplifying culture medium composition, wherein the activatingculture medium composition comprises an activating basal culture medium, rhIL-2, rhIL-15, rhIL-21, OK432, OKT-3 and Anti-CD16mAb or Anti-CD52mAb; the amplifying culture medium composition comprises anamplifying basal culture medium, rhIL-2, rhIL-15, rhIL-21 and DST. Through the adoption of the technology provided by the invention, the in-vitro preparation efficiency of NK is improved and the preparation cost is reduced; the NK purity of a final product is increased, and the obtained NK cells have higher biological activity.

The invention discloses a method for constructing tissue engineering skin, which comprises the steps as follows: (1) a tissue engineering skin nutrient solution is prepared; (2) a demic mechanocyte feeder layer is prepared; (3) a gelatin biomaterial is prepared; (4) the tissue engineering skin is constructed and cultivated. The method for constructing the tissue engineering skin adopts the adding of the mechanocytes after proliferation inhibition treatment to complete the interaction with epidermic cells so as to promote the mature and differentiation degree of the epidermic cells, has significant effect on improving the accuracy of compound toxicity detection in vitro to ensure that the prepared tissue engineering skin can more effectively resist the toxic action of the compound on competent cells and the forecast precision of the prepared tissue engineering skin to the compound toxicity is improved so as to create more effective animal substitution detection methods which are based on the tissue engineering skin.

The invention relates to methods for culturing human embryonic stem cells by culturing the stem cells in an environment essentially free of mammalian fetal serum and in a stem cell culture medium including amino acids, vitamins, salts, minerals, transferrin, insulin, albumin, and a fibroblast growth factor that is supplied from a source other than just a feeder layer the medium. Also disclosed are compositions capable of supporting the culture and proliferation of human embryonic stem cells without the need for feeder cells or for exposure of the medium to feeder cells.