135 results about "Feeder Layer" patented technology

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.

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.

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.

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 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 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.

The invention discloses a method for in-vitro separation and culture of goat male germ stem cells. Spermatogonia are obtained from the testicular convolutedtubules of a male goat. A method for separating and purifying the goat male germ stem cells adopts 0.2% of gelatin and a Matrigel differential adherence combined cloning method to differentiate and separate un-adhered goat male germ stem cellsfrom other cells for adherence culture; and the culture of the separated goat male germ stem cells is MEF feeder-layer culture of feeder-layer-free culture. The separated goat male germ stem cells have ES cell characteristics and the potential of differentiation into nerve cells, cardiac myocytes and sperm sample cells.