51 results about "Mesenchyme" patented technology

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The present application provides methods and processes for making and using a mesenchymal stem cell secretome for treating limbal stem cell deficiency (LSCD).

The invention belongs to the technical field of nano preparations, and particularly relates to a fused nano vesicle as well as a preparation method and application thereof. The fusion nano vesicle comprises a membrane structure formed by fusing a mesenchymal cell vesicle and a plant exosome, the mesenchymal cell vesicles are human umbilical cord mesenchymal cell vesicles; the plant exosome is an aloe exosome. Wherein the protein content ratio of the plant exosome to the mesenchymal cell vesicles in the nano vesicles is (1-20): (1-10). The fused nano-vesicle has the plant source advantages of low immunogenicity and high antioxidation and the human stem cell exosome advantages of strong immunoregulation and regeneration promotion, and has great potential in the aspects of tissue repair, anti-inflammatory treatment, drug delivery and the like.

The present invention provides a method of producing a population of mesenchymal stromal cells (MSC) of CD73 + CD44 +, CD90 +. The CD73 + CD44 +, CD90 + MSC is used in a method for producing terminally differentiated osteoblasts, adipoblasts and chondroblasts from pluripotent stem cells (PSCs). The differentiation methods include the use of a single agent-a WNT signaling pathway activator used on an adherent culture of PSC, such as a GSK3 [beta] inhibitor.

The application provides a WNT5B-promoting tooth differentiation biological product and an application thereof. Compared with the prior art, the application has the following advantages: the patent integrates human embryonic tooth germ spatial transcriptome sequencing and epithelial cell and interstitial cell extracellular protein group sequencing data, screens signal molecules WNT5B and CTNNB1 secreted in the development process of tooth germ epithelium, and confirms the important role of WNT5B protein in tooth germ development for the first time. Compared with other WNT family members, WNT5B has stronger tooth-forming ability of promoting tooth-derived stem cells; the effective concentration of WNT5B is low, and 10 ng / ml is the optimal concentration of WNT5B for promoting tooth-derived stem cells to form teeth; and the role of WNT5B and CTNNB1 in extracellular secretion of tooth germ epithelial cells and the synergistic effect of WNT5B and CTNNB1 in promoting tooth differentiation of tooth germ mesenchymal cells are further determined, that is, the combined low-concentration use effect of the two is better than the single high-concentration use.

The application relates to an IL-10 high-expression mesenchymal stem cell secretion component, a preparation method thereof and application thereof in prevention and treatment of ischemic cerebral stroke, and belongs to the technical field of cerebral stroke treatment. In order to solve the problems existing in the current stem cell clinical medication and administration method for ischemic cerebral stroke, the application provides a specially treated mesenchymal stem cell secretion component (hMSC-M), the secretion component is an IL-10 high-expression stem cell secretion component obtained by using a basic culture medium containing human IFN-gamma protein and TNF-alpha simulation peptide to induce human mesenchymal cells after expansion, and then harvesting culture and repeatedly freezing and thawing. The application finds that the hMSC-M has a significant effect on prevention and treatment of ischemic cerebral stroke through animal experiments, thereby opening up a new drug use for stem cell application, laying a foundation for developing high-efficiency related drugs for preventing and treating ischemic cerebral stroke injury, and providing a new application scheme and thought.

The invention provides a WNT5B dental differentiation promoting biological product and application thereof. Compared with the prior art, the application has the following advantages: by integrating human embryo tooth embryo space transcriptome sequencing data and epithelial cell and interstitial cell exocrine proteome sequencing data, signal molecules WNT5B and CTNNB1 secreted in the tooth embryo epithelial development process are screened out, the important effect of the WNT5B protein in tooth embryo development is confirmed for the first time, and the application has a wide application prospect. Compared with other WNT family members, the WNT5B has higher capability of promoting the tooth formation of the odontogenic stem cells; the onset concentration of the WNT5B is low, and the onset concentration is the optimal concentration of the WNT5B for promoting the odontogenic stem cells to form teeth when the onset concentration is 10ng / ml; the effects of the WNT5B and the CTNNB1 in tooth embryo epithelial cell exocrine and the synergistic effect of the WNT5B and the CTNNB1 in promoting tooth embryo mesenchymal cell odontogenic differentiation are further determined, namely the combined low-concentration use effect of the WNT5B and the CTNNB1 is superior to that of single high-concentration use of the WNT5B and the CTNNB1.

The invention provides application of an immune response biomarker in prognosis evaluation of mesenchymal stromal cells in treatment of liver cirrhosis patients, and relates to the technical field of biomedicine, the immune response biomarker comprises MX1 positive monocytes or / and LGALS2 positive monocytes, and the MX1 positive monocytes are MX1 positive monocytes or / and LGALS2 positive monocytes. Screening to obtain a potential biomarker LGALS2 positive mononuclear cell and an MX1 positive mononuclear cell for predicting the MSC treatment dose-effect relationship; compared with a healthy control, the proportion of baseline MX1 positive monocytes of a liver cirrhosis patient is remarkably increased (Plt; 0.01), while the proportion of LGALS2 positive monocytes is significantly reduced (Plt; 0.05) of the substrate (1); the change trend of the biomarker under the same MSC dose and the regulation effect on other immune cell subgroups are identified through sequencing analysis, the dose-effect relationship of MSC-mediated immune regulation can be accurately reflected, and patients with low MSC treatment prognosis recurrence risk can be identified.

An induction medium for increasing the IL-10 secretion of mesenchymal stem cells and its application, belonging to the technical field of stem cell culture. In order to provide a method that can both increase the IL-10 secretion of mesenchymal stem cells and not affect the proliferation of mesenchymal stem cells, the present invention uses a basal medium containing human IFN-γ protein and TNF-α mimetic peptide to induce the amplified human mesenchymal cells, and then after harvesting and culturing, mesenchymal stem cells with increased IL-10 secretion are obtained. The induction medium provided by the present invention can increase the IL-10 secretion of mesenchymal stem cells while not affecting the proliferation of mesenchymal stem cells, and the induced mesenchymal stem cells are expected to be applied to the preparation of cell drugs for treating inflammatory diseases.

To clarify the heterogeneous composition of mesenchymal stromal / stem cells (MSCs) and identify functional subpopulations of MSCs.SOLUTION: Provided is a method for identifying functional cell subpopulations in a cell population of MSCs, comprising: performing omics analysis of the cell population of MSCs at the single cell level; and clustering the cell population of MSCs based on the results of the analysis to classify it into cell subpopulations.SELECTED DRAWING: None

The present application provides methods and processes for making and using a mesenchymal stem cell secretome for treating limbal stem cell deficiency (LSCD).

Provided herein are methods of using mesenchymal stromal / stem cell (MCS) exosomes in the treatment of diseases associated with thymic dysfunction. In some embodiments, the MSC exosomes restore thymic architecture and development in a subject having thymic dysfunction (e.g., caused by exposure to hyperoxia). In some embodiments, the subject is a human subject (e.g., a human neonate).

The invention relates to methods for extracting, purifying and concentrating mesenchymal stromal cell exosomes enriched with micro-RNA, proteins and lipids, and can be used in cosmetology and pharmacology for rehabilitation and regeneration. The technical result of the invention is a more efficient process for producing exosomes. This technical result is achieved in that the present method for extracting, purifying and concentrating exosomes from mesenchymal stromal cells includes cultivating cells, collecting the conditioned culture medium, depleting high-molecular-weight proteins, extracting, purifying and concentrating exosomes, and freezing and lyophilizing the exosome concentrate; in order to enrich the culture medium with exosomes, multipotent mesenchymal stem cells (MSCs) are grown in a medium having a lowered glucose concentration and containing platelet lysate and 5 mM of L-alanyl-L-glutamine, wherein, in the case of static cell cultivation, the growth medium is placed in a culture vessel and once a level of not less than 80% monolayer confluence is reached, the culture medium is harvested and replaced once every 24 hours, and in the case of flow-through cell cultivation, cells are placed on cell carriers in a bioreactor and 72-96 hours after the start of cultivation, the culture medium is harvested and replaced once every 24 hours, the exosome-enriched culture medium is subjected to concentration, which includes successive stages of centrifugation, tangential flow filtration and isopycnic centrifugation: in the first stage, the culture medium is centrifuged for 10 mins at 2000 g to remove large cell particles and other inclusions; in the tangential flow filtration stage, the supernatant is concentrated using hollow fibre filters having a molecular weight cutoff of 500 kDa, after which the concentrate is subjected to fivefold diafiltration with an equal volume of DPBS phosphate buffer solution using the same hollow fibre cartridge; and in the isopycnic centrifugation stage, the concentrated supernatant is mixed with a saccharose solution and deuterium oxide and subjected to isopycnic centrifugation at 100000 g and 4˚С for 75 mins, after which the concentrate is subjected to further fivefold diafiltration with a buffer solution, then the concentrate is filtered through filters with a pore diameter of 0.22 µm and 0.1 µm, the exosome concentrate is subjected to initial freezing to -75˚C in a freezing chamber, and drying is carried out in a lyophilization chamber, wherein in a first drying stage, the concentrate is cooled to -40˚С and subsequently held for 100 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -30˚С and subsequently held for 610 mins at an absolute pressure in the chamber of not more than 10 Pa, then cooled to -10˚С and subsequently held for 315 mins at an absolute pressure in the chamber of not more than 10 Pa, after which it is cooled to 0˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa, then heated to +10˚С and subsequently held for 130 mins at an absolute pressure in the chamber of not more than 10 Pa; in a second drying stage, drying is carried out at +22˚С for not less than 130 minutes at an absolute pressure in the chamber of not more than 10 Pa.

A method of promoting return to estrus in an animal and an agent for promoting return to estrus in an animal are provided. The method is a method of promoting return to estrus in an animal, characterized by including a step of administering mesenchymal stromal cells or a composition containing mesenchymal stromal cells to the animal. The mesenchymal stromal cells are preferably adipose-derived stem cells, and the animal is preferably a farm animal, a dog, or a cat.

In certain aspects, the present invention provides methods for inducing a stable gene modification of a target nucleic acid via homologous recombination in a primary cell, such as a primary blood cell and / or a primary mesenchymal cell. In certain other aspects, the present invention provides methods for enriching a population of genetically modified primary cells having targeted integration at a target nucleic acid. The methods of the present invention rely on the introduction of a DNA nuclease such as a Cas polypeptide and a homologous donor adeno-associated viral (AAV) vector into the primary cell to mediate targeted integration of the target nucleic acid. Also provided herein are methods for preventing or treating a disease in a subject in need thereof by administering to the subject any of the genetically modified primary cells or pharmaceutical compositions described herein to prevent the disease or ameliorate one or more symptoms of the disease.

The invention discloses a human adenovirus species C having a capsid which comprises a modified adenovirus hexon protein, wherein the modified adenovirus hexon protein has a modified HVR1 region, wherein the modified HVR1 region has the sequence DEAATALEINLKKKKQAEQQ (SEQ ID NO.: 1). The invention further discloses the adenovirus of the disclosure for use in treating or preventing a human disease. The invention further discloses a nucleic acid encoding the modified adenovirus hexon protein. The invention further discloses the use of an adenovirus according to the disclosure for transducing mesenchymal stromal cells (MSCs) or tumor cells. The invention further discloses an in vitro method for transducing MSCs and a transduced MSC obtainable by the method. The invention further discloses the transduced MSC of the disclosure for use in treating a disease.

The invention discloses a large-scale stem cell culture method, and relates to the technical field of stem cell culture, and the method comprises the following steps: stem cell pretreatment: taking P3-P5 generation mesenchymal or embryo dry and thin parts, digesting for 2-3 minutes at 37 + / -0.5 DEG C by using trypsin containing 0.01%-0.05% of EDTA (Ethylene Diamine Tetraacetic Acid), the pretreated cells are inoculated to a bioreactor containing a modified microcarrier, the microcarrier is a chitosan and gelatin composite material, the aperture is 50-100 microns, the particle size is 200-300 microns, RGD peptide is grafted, dynamic fed-batch culture is conducted, the concentration of glucose and lactic acid is detected in real time through an intelligent feedback system, harvesting and purification are conducted, 0.02%-0.04% of collagenase is added after culture is finished, incubation is conducted for 15-20 min at the temperature of 37 DEG C, and the concentration of glucose and lactic acid in the bioreactor is detected. The limitation of a traditional single-material microcarrier is broken through, the chitosan and gelatin composite carrier is adopted, RGD peptide is grafted, the cell attachment rate is increased, a three-dimensional growth space is provided for cells through specific aperture / particle size design, and the problem of poor cell uniformity in large-scale culture is solved.

The present invention pertains to an improved mesenchymal stromal cell (MSC) preparation and a method for producing the same. The invention provides a new strategy to isolate MSC from bone marrow mononuclear cells (BM-MNCs) by pooling BM-MNCs of multiple unrelated (third-party) bone marrow donors. The MSC preparation manufactured in accordance with the methodology of the invention is characterized by a stable proliferative capability and an increased immunosuppressive potential when compared to individual donor MSC preparations or a pool of individual MSCs generated from multiple donors. The MSCs prepared according to the invention are particularly useful for medical applications such as the treatment of graft-versus-host disease (GvHD) in recipients with hematopoietic stem cell transplants, patients with autoimmune disorders and as a cell-based therapy in regenerative medicine.

The present invention relates to an inducer for transforming mesenchymal cells into epithelial cells, and the pyrrolopyridine derivative is a compound for inducing the transformation of mesenchymal cells into epithelial cells, and is represented by the following formula: JPEG2026500029000078.jpg52143 having formula (I) (wherein m1, m2, A2, A3 are described herein).

The present invention relates to the technical field of cell therapy. Specifically disclosed are a mesenchymal stromal cell-based drug and a use thereof in the treatment of acute cerebral infarction. The present invention provides a cell-based drug, comprising mesenchymal stromal cells and a pharmaceutically acceptable excipient. Compared with stem cells, the mesenchymal stromal cells have a negative expression level of mRNA encoding self-renewal and differentiation in the mesenchymal stromal cells. The mesenchymal stromal cells account for 90% or more of the cell-based drug. The cell-based drug exhibits the characteristics and mechanism of action of stromal cells, homes to the neovasculature formed by endothelial cells at the injury site within an effective time window, plays a role in promoting / maintaining the neovasculature and remodeling the microenvironment of the injury site, fully ensures the safety and effectiveness of disease treatment, and provides a novel method for the diagnosis and treatment of related diseases.

The invention discloses a preparation method and application of renal progenitor cells. The method specifically comprises the following steps: inducing pluripotent stem cells to be differentiated into an intermediate mesoderm cell stage through a GSK-3alpha / beta inhibitor, and inducing cells in the intermediate mesoderm cell stage to be differentiated into a cap-like mesenchymal stage by using a nuclear receptor agonist containing FGF-2 and RAR / RXR, so as to obtain the renal progenitor cells with obvious specific characteristics and potentials of the cap-like mesenchymal. According to the method, the high-purity and high-quality renal progenitor cells can be obtained without separation and purification, the purity can reach 99.9%, and the obtained renal progenitor cells have the differentiation potential of kidney-like organs, basically have no PSCs residues, have the differentiation potential of kidney-like organs and are suitable for clinical cell therapy; and the operation is simple, stable and controllable, and safe, effective, high-purity and homogeneous renal progenitor cells can be prepared on a large scale.

Provided is a method of producing a somite cell from a pluripotent stem cell, comprising the step of culturing a pluripotent stem cell in a medium comprising a GSK3β inhibitor. Provided is a method of producing a dermatome cell from a somite cell, comprising the step of culturing a somite cell in a medium comprising a GSK3β inhibitor and BMP. Provided is a method of producing a syndetome cell from a sclerotome cell, comprising the steps of culturing a sclerotome cell in a medium comprising FGF and then culturing the cell in a medium comprising BMP and TGFβ. Provided is a method of producing a mesenchymal stromal cell from a somite cell, comprising the step of culturing a somite cell in a medium comprising FGF. Provided are methods of producing a myotome cell, a dermatome cell, a sclerotome cell, and a syndetome cell from a pluripotent stem cell through a somite cell by appropriately combining the above methods and known methods.

This method for producing mesenchymal cells of the ventral hindgut mesoderm comprises: culturing pluripotent stem cells in an induction medium A containing a TGF-β inhibitor, a Wnt agonist, and a fibroblast growth factor to induce differentiation into epiblast-like cells; culturing the epiblast-like cells in an induction medium B containing a Wnt agonist, a fibroblast growth factor, and retinoic acid to induce differentiation into caudal epiblast-like cells; culturing the caudal epiblast-like cells in an induction medium C containing a Wnt agonist, a bone morphogenetic protein, and a TGF-β family member to induce differentiation into posterior primitive streak cells; and culturing the posterior primitive streak cells in an induction medium D containing a Wnt agonist, a fibroblast growth factor, a bone morphogenetic protein, and a hedgehog signaling agonist to induce differentiation into mesenchymal cells of the ventral hindgut mesoderm.

The application provides a nucleic acid molecule, a pro-angiogenic mesenchymal stromal cell and application thereof, relates to the biomedical technical field, and the nucleic acid molecule encodes HGF protein, or encodes VEGF165 protein and HGF protein; a first signal peptide is used as the signal peptide of the VEGF165 protein, and a second signal peptide is used as the signal peptide of the HGF protein. Through a gene modification strategy, MSC cells containing the nucleic acid molecule can stably and efficiently express VEGF165 and / or HGF, and the expression amount is significantly higher than that of MSC without the nucleic acid molecule. The cells have the ability to promote the migration of endothelial cells; meanwhile, the cells can also significantly promote the proliferation and tube formation of HUVEC cells, and further confirm the strong pro-angiogenic effect. The technical problem that nucleic acid molecules expressing VEGF165 or / HGF cannot be stably and efficiently expressed in the prior art is solved.

The invention relates to the technical field of biology, in particular to a preparation method of mesenchymal-like stromal cells derived on the basis of cat embryonic cell spheres. According to the preparation method of the mesenchymal-like stromal cells derived on the basis of the cat embryonic cell spheres, disclosed by the invention, the cell spherical aggregate is firstly cultured, and then adherent culture and induction are performed, so that the prepared cat mesenchymal-like stromal cells are higher in quality. In addition, by optimizing the formula of the induction culture solution, the concentration of the bFGF is improved, and the cell activity and the cell quality are effectively improved.

The invention relates to the technical field of organoid culture, and provides a mouse tooth embryo organoid culture method which comprises the following steps: step 1, taking a first molar tooth embryo of the lower jaw of an SD (Sprague Dawley) rat of which the embryo is 18.5 days; 2, dissociating the tooth embryo tissue into single cells by adopting a mechanical shearing and compound enzyme digestion method, wherein the compound enzyme is prepared by mixing collagenase I, neutral protease II and deoxyribonuclease I according to a mass ratio of 2: 2: 1; step 3, embedding the single-cell suspension in matrigel at a concentration of 150000 cells / 50 [mu] L; and 4, adding a DMEM / F-12 culture medium containing growth factors, and culturing for 7 days under the conditions of 37 DEG C and 5% CO2. The cultured organoid forms a layered structure similar to a clock-shaped tooth embryo on the seventh day, contains epithelial cells (CK14 positive) and mesenchymal cells (Vimentin positive), can express an enameblast marker AMELX, an odontoblast marker DSPP and a tooth development key factor TGF-beta, and is highly consistent with the in-vivo tooth embryo development characteristic.

Compositions and methods of transplanting cells by grafting strategies into solid organs (especially internal organs) are provided. These methods and compositions can be used to repair diseased organs or to establish models of disease states in experimental hosts. The method involves attachment onto the surface of a tissue or organ, a patch graft, a “bandaid-like” covering, containing epithelial cells with supporting early lineage stage mesenchymal cells. The cells are incorporated into soft gel-forming biomaterials prepared under serum-free, defined conditions comprised of nutrients, lipids, vitamins, and regulatory signals that collectively support stemness of the donor cells. The graft is covered with a biodegradable, biocompatible, bioresorbable backing used to affix the graft to the target site. The cells in the graft migrate into and throughout the tissue such that within a couple of weeks they are uniformly dispersed within the recipient (host) tissue. The mechanisms by which engraftment and integration of donor cells into the organ or tissue involve multiple membrane-associated and secreted forms of MMPs.

Provided are a method for producing a mesenchymal stem cell culture, a mesenchymal stem cell culture, a pharmaceutical composition containing a mesenchymal stem cell culture, and a method for producing a population of mesenchymal cells. This method for producing a mesenchymal stem cell culture includes a step (a) for culturing a tissue-derived sample containing a population of mesenchymal stem cells under serum starvation conditions or 2-deoxy-D-glucose (2-DG)-containing culture medium conditions to thereby separate a culture of mesenchymal stem cells that survived the serum starvation conditions or 2-DG-containing culture medium conditions. The population of mesenchymal stem cells is not exposed to growth conditions from after collection of the tissue-derived sample from an individual until culturing is performed under serum starvation conditions or 2-DG-containing culture medium conditions.

The invention provides a mesenchymal stromal cell (CiMS-IL10), which can be used for inducing fibroblasts to be reprogrammed into high-expression IL-10 (Interleukin-10) by virtue of a chemical small molecule combination, and an application of the CiMS-IL10 in the treatment of sepsis. The invention further provides a preparation method of the CiMS-IL10, which is used for inducing the fibroblasts to be reprogrammed into high-expression IL-10 (Interleukin-10) and an application of the CiMS-IL10 in the treatment of the sepsis and the application of the CiMS-IL10 in the treatment of the sepsis and the application of the CiMS-IL10 in the treatment of the sepsis. According to the invention, a chemical reprogramming strategy without genetic modification is adopted, and the fibroblasts from somatic cells are efficiently converted into CiMS-IL10 through sequential activation of an endogenous signal channel. The obtained cell can secrete IL10 at the post-reprogramming stage, and when the cell is applied to sepsis treatment, pathological symptoms can be improved and the death rate can be reduced by regulating inflammatory response. The invention provides a novel safe treatment strategy based on non-genetically modified cells for sepsis.