440 results about "Embryoid body" patented technology

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.

Methods are described for mapping a pathway of differentiation of a population of embryonic cells which includes exposing the cells to an exogenous factor and measuring gene expression products that are characteristic of a particular cell type or lineage. Directing differentiation of human embryonic cells relies on dissociated embryoid bodies which are then exposed to one or more exogenous factors to enrich a culture for a particular cell type. The differentiated cells may be used for treating a medical condition in a human. Kits for determining differentiation pathways and screening exogenous factors for their utility in differentiation are provided.

The present inventors identified aggregation of embryonic stem cells and embryoid bodies (EBs) as the cause of the difficulty in generating large numbers of the embryonic stem cells (ES) cell-derived tissues. To counter this, the invention provides a novel bioprocess where aggregation of spheroid forming cells, such as embryonic stem cells and spheroids, such as EBs is controlled, such as by encapsulation of within a matrix. As a result, EBs can be generated with high efficiency and cultured in high cell density, well-mixed systems. Well-mixed conditions facilitate measurement and control of the bulk media conditions and allow for the use of scalable bioreactor systems for clinical production of tissue. Therefore, the invention enables generation of ES cell-derived tissue on a clinical scale. The invention is also applicable to any spheroid-forming cells and other types of pluripotent cells.

The present invention relates to methods of using granulocyte colony stimulating factor (G-CSF) polypeptide, alone and in conjunction with stromal cell derived factor (SDF-1) polypeptide, to increase the mobilization of c-Kit+ stem cells in the blood, bone marrow, tissue, heart or other organs for the subsequent production of embryoid body-like cell clusters. These embryoid body-like cell clusters can be used for cell replacement therapy, for the treatment of cardiac myopathy and other diseases and disorders, and for screening agents that drive or inhibit differentiation and proliferation.

A method of generating cells predominantly displaying at least one characteristic associated with a cardiac phenotype is disclosed. The method comprises (a) partially dispersing a confluent cultured population of human stem cells, thereby generating a cell population including cell aggregates; (b) subjecting said cell aggregates to culturing conditions suitable for generating embryoid bodies; (c) subjecting said embryoid bodies to culturing conditions suitable for inducing cardiac lineage differentiation in at least a portion of the cells of said embryoid bodies, said culturing conditions suitable for inducing cardiac lineage differentiation including adherence of said embryoid bodies to a surface, and culture, medium supplemented with serum, thereby generating cells predominantly displaying at least one characteristic associated with a cardiac phenotype.

Primate embryoid bodies are formed from primate ES cells. The ES cells form clumps. One then removes the clumps, as clumps, and permits incubation under non-adherent conditions. The development of embryoid bodies from primate ES cells is dependent on maintaining the aggregation of cells, as individualized cells will rapidly die.

The invention relates to a submerged plant breeding blanket and application thereof. The submerged plant breeding blanket disclosed by the invention is respectively composed of a carrier layer, a matrix layer and an adhesive layer from bottom to top, wherein the carrier layer is composed of palm and bamboo plant fiber; the matrix layer is composed of a growing matrix suitable for submerged plants to grow; and the adhesive layer is composed of a plant fiber net. The invention further provides a submerged vegetation transplantation technology which is characterized in that after being subjected to seedling culture, submerged plant seedling breeding embryoid bodies prepared by virtue of the breeding blanket are transplanted to the water bottom of an ecological restoration area for planting and restoring submerged vegetation of the ecological restoration area. Compared with the traditional submerged plant transplantation technology method, the submerged vegetation transplantation technology disclosed by the invention has the advantages of low transplantation cost, high speed, high survival rate, convenient technology, strong technical maneuverability, high community forming speed after transplantation, high breeding speed, good community stability and the like, and can be used for realizing the quick transplantation and the community construction and configuration of the submerged plants.

A three-dimensional microwell system that supports long term pluripotent cell culture and formation of homogeneous embryoid bodies (EBs) is described. Microwell-cultured pluripotent cells remain viable and undifferentiated for several weeks in culture and maintain undifferentiated replication when passaged to Matrigel®-coated, tissue culture-treated polystyrene dishes. Microwell-cultured pluripotent cells maintain pluripotency, differentiating to each of the three embryonic germ layers. Pluripotent cell aggregates released from microwells can be passaged for undifferentiated replication or differentiated to monodisperse EBs. The ability to constrain pluripotent cell growth in three dimensions advantageously provides for more efficient, reproducible culture of undifferentiated cells, high-throughput screening, and the ability to direct pluripotent cell differentiation by generating monodisperse EBs of a desired size and shape. Cardiomyocyte-rich EBs are obtained from pluripotent cells cultured in microwells of defined size and shape.

Disclosed are embryoid bodies having a uniform size of approximately 415 nm and comprising genetically modified embryonic stem cells, and methods of making same. The genetically uniform embryoid bodies can be multiplexed as one embryoid body per well in a multiwell format, and used as a high to medium throughput screen for test agents that affect the development and homeostasis of animals, including humans. The genetic modification of the embryonic stem cells is a promoter-report-selection construct that enables the selection and detection of cells of a particular lineage in the EB, to determine the effects of a test agent.

A method is provided to prepare a plurality of microwells suitable for the formation of embryoid bodies from embryonic stem cells. The method requires applying an image-forming material to a heat sensitive thermoplastic material in a designed pattern and heating the material under conditions that reduce the size of the receptive material by at least about 60% to create a mold. A polymer such as PDMS is then applied to the mold and removed to form the microwells. In an alternative aspect, the plurality of microwells on receptive material are prepared by etching a microwell designed pattern into a heat sensitive thermoplastic material support and then heating the material under conditions that reduce the size of the receptive material by at least about 60%.

The invention relates to the field of stem cell biology, specifically to a method for differentiating human pluripotent stem cells into natural killer cells and an application. The invention disclosesa pluripotent stem cell-derived natural killer cell. The pluripotent stem cell-derived natural killer cell expresses CD56, Nkp30, Nkp44 and Nkp46, and also expresses markers CD16 and CD94 of a maturenatural killer cell. The invention also discloses a method for preparing the natural killer cell. The method comprises the following steps: S1, formation of an embryoid body; S2, differentiation of the embryoid body into hematopoietic progenitor cells; S3, differentiation of the hematopoietic progenitor cells into NK cells; and S4, maturation and expansion of the NK cells. With the differentiation method provided by the invention, the pluripotent stem cells can be rapidly, efficiently, simply and conveniently induced to be differentiated into the natural killer cells with low cost on the basis of a culture medium with definite components and an optimized cell factor combination.

Methods of tissue engineering, and more particularly methods and compositions for generating various vascularized 3D tissues, such as 3D vascularized embryoid bodies and organoids are described. Certain embodiments relate to a method of generating functional human tissue, the method comprising embedding an embryoid body or organoid in a tissue construct comprising a first vascular network and a second vascular network, each vascular network comprising one or more interconnected vascular channels; exposing the embryoid body or organoid to one or more biological agents, a biological agent gradient, a pressure, and/or an oxygen tension gradient, thereby inducing angiogenesis of capillary vessels to and/or from the embryoid body or organoid; and vascularizing the embryoid body or organoid, the capillary vessels connecting the first vascular network to the second vascular network, thereby creating a single vascular network and a perfusable tissue structure.

Culturing embryonic stem cells without the use of embryoid bodies leads to a increase in the frequency of predetermined cell types.

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1 and TWIST but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiated the stem cells, are also provided.

Alginate polyelectrolyte encapsulation is used for the controlled differentiation of embryonic stem cells. An isolated cell population is provided. The cell population includes a single cell suspension of ES cells encapsulated within an alginate polyelectrolyte microenvironment. The encapsulated ES cells are capable of differentiating within said microenvironment into hepatocyte lineage cells in the absence of embryoid body intermediates or growth factor supplementation.

The invention discloses a honeycomb SCR (Selective Catalytive Redution) smoke denitration catalyst embryoid body and a preparation method thereof. The honeycomb SCR smoke denitration catalyst body consists of the following raw materials as per percentage by weight: 40 to 60 percent of titanium dioxide, 1 to 10 percent of white clay, 0.1 to 1 percent of ammonium metavanadate, 1 to 8 percent of ammonium meta-tungstate, 0.1 to 3 percent of niobium oxalate, 0.5 to 3 percent of lactic acid aqueous solution, 1 to 5 percent of oxalic acid aqueous solution, 25 to 35 percent of deionized water, 1 to 10 percent of ceramic fiber, 0.1 to 3 percent of fluff pulp, 0.1 to 1 percent of stearic acid, 0.1 to 1 percent of hydroxypropyl methyl cellulose and 1 to 5 percent of glycerol. The honeycomb SCR smokedenitration catalyst embryoid body has high plasticity, caking property and mechanical strength. The method is simple and convenient in operation and low in cost. Each component of the prepared catalyst body is dispersed uniformly; the catalyst body is easy in forming and demolding in the extrusion process and has high finished product rate; and the catalyst prepared by the embryoid body has highdenitration efficiency.

The present invention provides a method of generating megakaryocytes and platelets. In various embodiments, method involves the use of human embryonic stem cell derived hemangioblasts for differentiation into megakaryocytes and platelets under serum and stromal-free condition. In this system, hESCs are directed towards megakaryocytes through embryoid body formation and hemangioblast differentiation. Further provided is a method of treating a subject in need of platelet transfusion.

Production of beta-cells from stem cells from pluripotent stem cells have always been significantly lacking in at least one of the following properties: 1) functional properties related to insulin-production and glucose signaling response, 2) mature phenotype such as biochemical markers or cell structures, 3) efficiency in production of differentiated cells. Described herein is multistep differentiation protocol which substantially overcomes all of the existing limitations. Pluripotent stem cells, including induced pluripotent stem cells (iPSCs), and embryonic stem cells (ESCs) can be differentiated using an embryoid body (EB) formation step, followed by B maturation via endothelial cells (EC) co-culturing and incubation with a sequential series of bone morphogenic protein (BMP)-related growth factor cocktails. The resulting cells displayed functional properties, including insulin-production and glucose signaling response, and mature phenotype of C-peptide expression.

The invention discloses a method for building a high-efficiency regeneration system of superior corn self-bred line agriculture line 531, belonging to the field of plant genetic engineering and transgenosis breeding. The invention takes an agriculture line 531 rataria as an explant, induces in a callus induction medium and produces an II-type embryonic callus; the II-type embryonic callus is subjected to embryoid induction under light in an embryoid induction medium to produce a green embryoid; then, the green embryoid is transported to a regeneration medium and is cultured into a regeneration plant under light; root induction is carried out in a rooting medium, and acclimatization is carried out in Hogland nutrient solution to ensure that a new thick root grows on the root of the regeneration plant; the root is transplanted to nutritional soil for rejuvenation culture; and finally, the root is transplanted to a land for growing field crops to normally grow and seed. The regeneration technology is suitable for the superior corn self-bred line agriculture line 531 with high application value, can ensure that the superior quality of agriculture line 531 corn can be inherited in the corn transgenosis breeding process, and has an important meaning for functional genome group research.

The invention provides a method for obtaining doubled haploid (DH) plants of sweet peppers, which comprises the following steps of: (1) selecting buds of a donor plant, the microspores of which are in a development state between nonokaryotic stage with the nucleus located aside and early dikaryotic stage, and pretreating at 4 DEG C for 1-3 days; (2) stripping the calyces off, soaking in alcohol, disinfecting through oscillation with sodium hypochlorite, and cleaning with sterile water; (3) stripping the anthers, and inoculating the anthers on an N4-3 solid-liquid double-layer culture medium by the density that 12-18 anthers are inoculated to a culture dish with diameter of 60 mm; (4) firstly, culturing the anthers for 1-10 days in the dark at 28-35 DEG C, and then, transferring to the condition of 25-28 DEG C to continue culturing in the dark; and (5) culturing for 4-7 weeks, when a large quantity of embryoids occur, transferring leaf type embryoids to an MS basic culture medium free of hormones, and cultivating the seedlings. According to the cultivating method provided by the invention, the occurrence rate of the embryoids breakthrough the limitations of genotype, the culturing efficiency of sweet peppers is greatly increased, and meanwhile, the ratio of the DH plants is also increased.

The invention relates to a vessel for embryoid formation used for forming embryoid bodies from ES cells easily without complicated technique, and to a method for forming embryoid bodies easily and efficiently using the vessel. The method includes the steps of (A) providing a vessel for embryoid formation having a coating layer formed from a compound having a particular PC-like group on a vessel surface defining a region for floating culture of ES cells, and (B) floating culturing ES cells in the vessel to form embryoid bodies.

The invention provides a high active induction culture method for lily bulbil which is characterized in that: through an induction culture of embryoid and tissue enrichment culture of embryoid tissue, a relative high rapid propagation velocity is obtained; then through an differentiation culture of bulb-lets and an intumescence culture of bulb-lets, transplanting bulb-lets of a circumferential diameter bigger than 3cm are obtained. The growth rate of the bulb-lets of the invention can reach more than one million grains per years which is ten times that of conventional methods. With a large number of small bulb-lets obtained, the ratio of bulb-lets with large specification and large grain size is increased. The invention optimizes the culture procedures of preferential lily seedball, shortens the propagation time of preferential lily seedball; furthermore, the field planting survival rate of bulb-lets is high and the production cost is low; the invention is suitable for the scale rapid propagation production of cut lily elite.