23 results about "Cell differentation" patented technology

The present invention discloses an application of highly active human umbilical cord mesenchymal stem cells. HA-MSCs were isolated from human umbilical cord tissue using TeSR-E8 containing 5% CloneR and LN521-Coated culture flasks, and a technical system for HA-MSC isolation, expansion, and cryopreservation was established. Compared with conventionally cultured MSCs, the HA-MSCs prepared by the present invention have higher proliferation activity, smaller size, a higher nuclear-cytoplasmic ratio, and highly express embryonic stem cell-associated marker antigens SOX2, Nanog, and OCT4. They have the potential to differentiate into neural, myocardial, and hepato-intestinal progenitor cells derived from the three germ layers and possess excellent anti-aging activity.

The invention relates to the technical field of cell differentiation, in particular to a method for directionally differentiating iPSC into iNK. Specifically, the method for preparing the CD34 + hematopoietic stem / progenitor cell and the iNK on the shaking table comprises the following steps: forming an embryoid (EB) by the iPSC on the shaking table, and then collecting a single cell supernatant to obtain the iNK. According to the method, high-proportion CD34 < + > hematopoietic stem / progenitor cells are obtained by adjusting the factor concentration and collecting the supernatant and are further induced and differentiated into the iNK, and more than 100 iNKs can be harvested before amplification of one iPSC according to the iNK harvested in batches.

Provided is a method for obtaining the germ cell lineage of an oviparous vertebrate more efficiently than conventional techniques. A host oviparous vertebrate is prepared at a developmental stage after the development of black pigmentary cells in the retina and before the formation of multiple layers of germ cells in the genitals, and isolated undifferentiated germ cells from a donor oviparous vertebrate are transplanted into the host oviparous vertebrate.

The current disclosure provides methods and compositions that provide for the insertion of transgenes in stem or progenitor cells without the deleterious effects to T cell differentiation by in vitro T cell differentiation methods. To accomplish this, the inventors have discovered that expression of the transgenes under the control of promoter regions, such as CD8a or CD8b, allows for the coordinated expression pattern that provides for both: 1) high expression of the transgene in mature T cells and 2) a coordinated level of expression of the transgene throughout the in vitro differentiation method that allows for the production of a population of mature T cells.

The invention relates to the technical field of cell differentiation and the technical field of myocardial tissue regeneration medicine, in particular to an induction culture medium and a method for rapidly obtaining myocardial balls capable of jumping spontaneously, hiPSC-sourced myocardial cells are obtained through co-culture with hMSC, and a differentiation culture medium is combined, so that myocardial cell clones harvested by D7 are more independent, and the differentiation efficiency is improved. The myocardial cells are naturally separated from other non-myocardial cells and can be rapidly cloned, screened and purified by using physical and enzyme-free reagents, so that the step of screening and purifying a culture medium in a traditional method is omitted, and the time and differentiation cost are saved.

The present invention provides a method for producing a natural killer cell from an iPS (Induced pluripotent stem) cell, comprising steps of: (i) contacting an iPS cell with a composition comprising a GSK-3 inhibitor and a ROCK inhibitor to obtain an embryoid body, (ii) contacting the embryoid body with a composition comprising a TGFβ receptor inhibitor to obtain a hematopoietic progenitor cell, (iii) culturing the hematopoietic progenitor cell to obtain a lymphocyte progenitor cell, and (iv) differentiating and expanding the lymphocyte progenitor cells to a natural killer cell.

The invention discloses a bovine liver organoid culture medium and a bovine liver three-dimensional organoid culture method, and belongs to the technical field of cell culture. The bovine liver organoid culture medium comprises a bovine liver cell amplification culture medium and a bovine liver cell differentiation amplification culture medium. The three-dimensional organoid culture method comprises the following steps: dissociating bovine liver tissues by using a liver dissociation solution, and then incubating by using a red blood cell lysis buffer solution to obtain primary liver cells; preparing a bovine liver stem cell suspension from matrigel and the primary liver cells, planking, performing inverted culture to enable the matrigel to form a dome shape, performing upright culture, and sequentially performing multiplication culture through a bovine liver cell amplification culture medium and differentiation culture through a bovine liver cell differentiation amplification culture medium. According to the method disclosed by the invention, efficient acquisition, amplification and directional differentiation of the bovine liver primary cells are realized by customizing the culture medium, optimizing the cell separation process and adopting a 3D culture technology, and the blank of a large economic animal disease research model is filled.

The invention relates to the field of cell differentiation and cryopreservation, in particular of pancreatic lineage cells. It provides methods for differentiating cells of the pancreatic lineage, in particular to islet-like clusters. It further provides methods for freezing cells of the pancreatic lineage, in particular endocrine progenitor cells. It also provides new pancreatic lineage cell populations.

The invention discloses a preparation method of an embryoid body for overexpressing PRDM1, which accelerates the formation and maturation process of the embryoid body and shortens the amplification period of the embryoid body through a lentivirus-mediated PRDM1 overexpression system, reveals the regulation mechanism of the embryoid body in combination with a genomics technology, and provides a new technical strategy and theoretical basis for optimizing the differentiation efficiency of stem cells.

An in vitro method of producing a population of cells comprising the steps of introducing a source of cells into the first cell culture chamber of an in vitro cell culture vessel comprising three cell culture chambers arranged in a series, each chamber being separated from the next chamber in the series by a semipermeable barrier; contacting the cells in the first cell culture chamber to a first composition of one or more cell growth factors, cell activating factors, cell differentiation factors, cytokines and cell attractants so as to promote migration of at least a subset of cells from the first cell culture chamber into an intermediate cell culture chamber of the series; and contacting cells in the intermediate cell culture chamber of the series to a further composition of one or more cell growth factors, cell activating factors, cell differentiation factors, cytokines and cell attractants so as to promote migration of at least a subset of cells from the intermediate cell culture chamber to the final cell culture chambers in the series. Also related cell culture vessels, cell populations and methods of using cell populations.

This invention provides a concentration gradient cell culture system that solves the problem of large errors in manual drug addition and the difficulty in constructing a stable, continuous concentration gradient in studies on the control of osteoclast progenitor cell differentiation using propionic acid-GPR43. [Solution] The system comprises a housing assembly with an inner container, a first liquid storage cylinder 2, a second liquid storage cylinder 3, a tuned crossbeam 4, an adjustable screw rod 5, a flow-dividing branch pipe array frame, a spiral mixing unit, a porous diffusion plate, and a culture tray 9. The tuned crossbeam is driven by the adjustable screw rod to synchronously supply liquid from both liquid storage cylinders. After gradient distribution by the flow-dividing branch pipe array frame and mixing by the spiral mixing unit, the porous diffusion plate forms a continuous and stable propionic acid concentration gradient field on the culture tray. This avoids manual errors, provides accurate concentration gradients to RAW264.7 and BMMs cells in a single experiment, and supports the calculation of dose-effect curves and IC50, as well as the elucidation of the mechanism of osteoclast differentiation control by propionic acid-GPR43 signaling.

This invention discloses a specific expression regulatory element for the hornless target gene RXFP2 in goats and its application. The sequence of the regulatory element is SEQ ID No. 1. Through single-cell gene expression and chromatin accessibility analysis of fetal bovine horn bud tissue, this invention discovered a specific, open regulatory element (chr12:29279059-29279559) upstream of RXFP2 in horn bud tissue. This element possesses a binding motif for RUNX2, a core transcription factor promoting osteoprogenitor cell differentiation, and is highly conserved in horned animals. Its location in the goat genome is chr12:57440948-57441449. Dual-luciferase assays verified that this regulatory element mediates the interaction between transcription factors RUNX2 and RXFP2, achieving specific high expression of RXFP2 in horn bud tissue. By knocking out the regulatory element region (chr12:57440800-57441678) in goat embryos using CRISPR-Cas9, goat embryos with gene-edited knockout of the hornless target gene RXFP2 specifically expressing regulatory elements were successfully prepared. This invention further elucidates the origin, evolution, and skin ossification mechanisms of horn organs, providing new targets for hornless livestock breeding.

The present invention discloses a medical gasket that does not shed fibers and a preparation method thereof. The medical gasket that does not shed fibers consists of a PTFE felt sheet in the middle layer and biodegradable electrospun film layers on the upper and lower surfaces as the main body; a laser drilling process is used to form uniformly distributed through holes on the main body of the medical gasket, and a layer of biodegradable biomaterial coating is covered on the surface of the through holes; the medical gasket is used in vascular surgery to achieve the effects of decompression and hemostasis; among them, the electrospun film layer is made of a biodegradable biomaterial, has good biocompatibility, not only effectively avoids the phenomenon of fluff shedding of the PTFE felt sheet, but also can guide and support cell differentiation and promote tissue regeneration; the through holes coated with the biodegradable biomaterial coating are beneficial to the insertion of the suture needle, improving the convenience of the operation.

The technology relates in part to methods for detecting the activity of IL-40 and modified versions thereof. The technology also relates in part to uses of IL-40 for promoting cell differentiation.

The present invention discloses SNP sites in the promoter region of the MMP2 gene and their application in pig breeding. The present invention takes the promoter region of the MMP2 gene as the research object, and uses molecular and cell biological methods to study its correlation with the sexual maturity of sows: it is found that the g.30088284G>A site and the g.30088004C>T site are significantly correlated with the age of sexual maturity of the Douro Black pig group; then, by transfecting different vectors, the different genotypes of the above sites are studied to affect the activity of the IRS1 gene promoter. Further, using sow ovarian granulosa cells as experimental materials, transcriptome sequencing, total iron colorimetry, malondialdehyde colorimetry and other experimental techniques are used to study the effect of the MMP2 gene on the transcription level and ferroptosis of sow ovarian granulosa cells. It is found that the differential genes in the cell transcriptome affected by the MMP2 gene are mainly enriched in signal pathways such as cell differentiation, promoting cell ferroptosis.

The present application relates to the technical field of cell differentiation, in particular to a method for differentiating iPSC into iNK. Specifically, the present application relates to a method for preparing CD34+ hematopoietic stem / progenitor cells and iNK on a shaker, which comprises the following steps: forming embryoid bodies (EB) of iPSC on a shaker, and then obtaining iNK by collecting single-cell supernatant. By adjusting the concentration of factors and the method of collecting supernatant, the present application can obtain a high proportion of CD34+ hematopoietic stem / progenitor cells and further induce the differentiation into iNK. The iNK obtained by batch harvesting can reach more than 100 iNK before the iPSC is expanded.

The utility model relates to the technical field of calculus bovis cultivation, and discloses a bioreactor for in-vitro cultivation of calculus bovis, which comprises a reactor body, a material uniformizing mechanism arranged in an inner cavity of the reactor body, a plurality of groups of oxygen supply tanks arranged at the upper end of the reactor body, and a first electromagnetic valve arranged at the lower end of each group of oxygen supply tank, an oxygen supply mechanism is arranged at the bottom of the inner cavity of the reactor body and is arranged below the refining mechanism; through cooperation of the oxygen supply mechanism, the reactor body and the material uniformizing mechanism, oxygen can be uniformly introduced into the bottom of the inner cavity of the reactor body through the oxygen supply mechanism, so that the contact range between the oxygen and cells is expanded, and meanwhile, under the action of the material uniformizing mechanism, the oxygen can flow in the reactor body along with bile, so that the bile is uniformly dispersed. Therefore, the oxygen can be uniformly supplied to the cells, and the mixing uniformity of the bile and various raw materials for promoting cell differentiation is improved, so that the cultivation efficiency of the calculus bovis is improved.

The invention discloses a system for promoting in-vitro differentiation of chicken oogonial stem cells and forming blastocysts, and belongs to the technical field of stem cell differentiation culture. According to the invention, retinoic acid (RA) and vitamin C (VC) are added on the basis of an oogonial stem cell culture medium to induce cells to start meiosis, and then follicle stimulating hormone (FSH), human chorionic gonadotropin (HCG), glial cell-derived neurotrophic factor (GDNF), progesterone (P4) and rosiglitazone are further added on the basis of the culture medium containing RA and VC to promote the recovery of meiosis, so that the effect of promoting meiosis is achieved. A complete system that the chicken oogonial stem cells are differentiated in vitro and form blastocysts is creatively established by inducing formation of secondary oocytes, direct evidence is provided for in-vitro reconstruction of poultry ovum generation, and a technical basis and application prospects are provided for preservation of female germ cells and large-scale production of the chicken oocytes.

Isolated populations of mesenchymal stem cells (MSCs) are provided, including mammalian mesenchymal stem cells (MSCs) characterized by the lack of CD54 (CD54−) or low cell-surface CD54 (CD54low). Methods of in vitro cell differentiation and methods of treatment using said isolated populations are also provided.

The invention discloses application of fructo-oligosaccharide in improvement of growth performance and intestinal development of suckling piglets and a nutrition regulator of the fructo-oligosaccharide, the fructo-oligosaccharide is applied to the suckling piglets as an early nutrition intervention means, the intestinal villus height of the suckling piglets is remarkably increased, the crypt depth is reduced, and the growth performance of the suckling piglets is improved. The improvement of the activity of small intestine disacidase and the change of small intestine stem cell differentiation marker gene mRNA indicates that the fructo-oligosaccharide improves the growth performance of the suckling piglets by promoting the intestinal tissue morphology, digestion and absorption functions and small intestine stem cell differentiation capacity of the suckling piglets.

The invention discloses a preparation method and application of a dendritic cell-derived exosome, and the preparation method comprises the following steps: (1) inducing in vitro differentiation of bone marrow-derived dendritic cells (DCs) to tolerant dendritic cells (tolDCs) through interleukin-10, and ensuring that the tolDCs highly express key antioxidant and anti-inflammatory proteins, namely, heme oxygenase-1 (HO-1); (2) separating and purifying an exosome HO-1 high-tolDex from the cell culture supernatant of the tolDCs (HO-1 high-tolDCs) with high expression of the HO-1 by virtue of methods such as ultracentrifugation and the like; (3) carrying out comprehensive physicochemical property and biological function identification on the prepared HO-1high-tolDex, and (4) verifying that the HO-1high-tolDex can effectively inhibit excessive activation and inflammatory response of target cells through an in-vitro cell experiment and an in-vivo experiment of a systemic lupus erythematosus (SLE) animal model, alleviate immune pathological injury of model animals, inflammation of kidneys and oxidative stress injury, and improve the bioavailability of the model animals. The immune tolerance is induced; the cell-free immunomodulator is a new generation of efficient and safe cell-free immunomodulator.

The invention discloses an adipogenic induced differentiation culture medium suitable for natural immortalized cells of chick embryos, an adipogenic differentiation method and application of the adipogenic induced differentiation culture medium and the adipogenic differentiation method, and belongs to the field of poultry cell differentiation. The chick embryo naturally immortalized cells are obtained by naturally immortalizing and screening 9-11-day-old SPF chick embryos, and auxiliary factors of the induced differentiation medium comprise dexamethasone, insulin, IBMX, rosiglitazone, oleic acid, glutamine, transferrin and BSA. The culture medium suitable for the natural immortalized cells of the chick embryos has a good adipogenic differentiation effect, almost all cells have lipid droplets after the natural immortalized cells of the chick embryos are subjected to in-vitro induced differentiation for 48 hours, the cell differentiation efficiency reaches about 95% or above, the differentiation efficiency of the natural immortalized cells of the chick embryos to an adipocyte lineage is remarkably improved, and the differentiation rate of the natural immortalized cells of the chick embryos to the adipocyte lineage is greatly improved. And the differentiation time is greatly shortened, so that the method has important significance on the research on the lipogenic differentiation mechanism of the fiber-derived cells and the preparation of cell culture meat.