93 results about "Conditioned medium" patented technology

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

The invention discloses stem cell freeze-dried powder for promoting skin healing and a preparation method of the stem cell freeze-dried powder. The freeze-dried powder comprises an effective component, a freeze-drying protective agent group and a polymer protective agent group, the effective component is umbilical cord mesenchymal stem cell conditioned medium freeze-dried supernatant, the protective agent group is selected from one or more of cane sugar, trehalose, lactose and mannitol, and the polymer protective agent group is selected from one or more of polyvinylpyrrolidone (PVP) and glucan. The preparation method comprises the following steps: collection of a conditioned culture medium, preparation of a freeze-drying protective agent solution, mixing treatment, low-temperature pre-freezing and freeze-drying, collection and packaging of freeze-dried powder and the like. The method can effectively retain stem cell-derived active factors and enhance the stability and redissolvability of the freeze-dried powder, and is suitable for skin wound repair, regenerative medicine and other scenes.

Provided herein are methods and related compositions for enhancing or enhanced partial reprogramming of target cells in a subject in need thereof (e.g., a human subject suffering from or at risk of a disease), the method comprising administering a plurality of mesenchymal lineage progenitor or stem cells (MLPSCs), exosomes derived therefrom, or conditioned culture media derived therefrom to a subject that expresses or will express one or more reprogramming factors in a population of target cells, whereby a plurality of the target cells in the subject become partially reprogrammed, but not fully reprogrammed.

Provided herein are cell-free compositions containing the secretome / exosomes of dsRNA primed cells ("herein dsRNA-primed secretome / exosome). The dsRNA-primed secretome is obtained from conditioned media following cell culture (for an effective amount of time) of cells treated with dsRNA, either by transfecting the cells with dsRNA or incubating the cells with dsRNA. The dsRNA-primed secretome / / exosome can be administered to a subject in need thereof, by administering an effective amount of the dsRNA-primed secretome / exosomes to a subject in need thereof. Exemplary subjects / sites of treatment include subjects include, but are not limited to in need of wound healing such as skin wound healing, burn sites, subjects in need of bone repair such as osteochondral bone repair, bone fracture sites, corneal wound sites. Other subjects include osteoporotic subjects and subjects with intervertebral disc degeneration.

The invention provides an aerosol-type face pack cosmetic that, when dispensed, forms a flexible, easy-to-use, and low-irritation resin foam that easily adheres to the face, such as around the eyes and nose. It also allows the fermented liquid contained in the resin foam to easily seep out, making it easy to apply the fermented liquid to the face and less likely to cause tightness. [Solution] An aerosol pack cosmetic comprising a beauty serum and dimethyl ether, wherein the beauty serum comprises at least one fermentation solution selected from Saccharomyces / Anzange ferment filtrate, Saccharomyces / Human (fibroblast / mononuclear cell) induced pluripotent cell culture conditioned medium ferment filtrate, Galactomyces / Eryngium multiflorum callus culture ferment filtrate, and a resin.

The invention relates to the technical field of biology, and discloses a construction method of a gastric cancer organoid culture system, which comprises the following steps: preparation of a conditioned medium: culturing L-WRN cells and collecting the conditioned medium; preparing a gastric cancer organoid culture medium, and mixing the conditioned culture medium, the basic culture medium, a plurality of growth factors and additives; and extraction and culture of organoid: separating cells from gastric cancer tissues, mixing the cells with matrigel, and culturing by using the prepared culture medium. According to the method, the repeatability and the stability of the organoid model are improved, the response consistency of the organoid in drug screening is improved, and the reliability of the model in disease mechanism research and personalized medical application is guaranteed.

The invention belongs to the technical field of biological medicines, and particularly relates to application of miR-206-3p in regulation and control of skeletal muscle atrophy, and the change condition of the miR-206-3p level is measured by constructing an in-vitro cancer cachexia myotubular atrophy model; it is verified that in-vitro addition of miR-206-3p mimics relieves in-vitro myotube atrophy induced by a cancer conditioned culture medium, inhibits expression of a myotube marker gene and inhibits proteolysis and metabolism pathways and cell apoptosis of myotubes, so that myotube atrophy induced by cancer cachexia is inhibited. The invention opens up a new thought for the research of cachexia pathogenesis, and also provides a potential new target for the research and development of cachexia treatment and / or cachexia prevention drugs.

The invention relates to a method for constructing a last-stage atherosclerotic plaque through additive manufacturing, and belongs to the technical field of tissue engineering. According to the method, firstly, cells are treated, then bio-ink for a fiber cap, bio-ink for a lipid nucleus and bio-ink for a suspension pool are prepared respectively, and then construction of the last-stage atherosclerotic plaque is completed through curing treatment. After a conditional culture solution and a stimulating factor are added, long-term in-vitro culture can be realized, and a real focus microenvironment can be simulated. The fibrous cap-lipid nucleus AS plaque constructed by the invention can maintain the stability and physiological function of the structure for a long time, and can accurately simulate hyperplasia, hypoxia, fat enrichment and inflammatory response of the AS plaque in the last stage.

The invention belongs to the technical field of cell biology, and particularly relates to a feeder-layer-free and serum-free bovine embryonic stem cell culture medium, a culture system and a culture method. The bovine embryonic stem cell culture medium comprises a conditioned culture medium and additives, the additives are IWR-1 and activin A, and the concentrations of the IWR-1 and the activin A in the conditioned culture medium are 2.5-5 [mu] M and 20-25 ng / mL respectively; the culture system comprises a bovine embryonic stem cell culture medium, gelatin with the mass concentration of 0.1-0.2% and a subculture medium, and the gelatin with the mass concentration of 0.1-0.2% is used for providing an attachment matrix for bovine embryonic stem cells; the subculture medium is prepared by adding Y27632 into a bovine embryonic stem cell culture medium. The cost of the extracellular matrix is reduced, and the feeder-layer-free bovine embryonic stem cell has high single cell passage efficiency and is convenient for gene editing operation.

The invention relates to the technical field of biological medicine, and provides application of a dental pulp stem cell exosome in preparation of a medicine for treating and preventing osteoarthritis, a preparation method of the dental pulp stem cell exosome comprises the following steps: S1, obtaining dental pulp tissue from a dental pulp cavity of an in-vitro tooth; s2, pretreating dental pulp tissues, adding digestive juice, digesting, centrifuging, discarding supernatant, adding a complete culture medium containing antibiotics into precipitates, mixing, filtering and centrifuging to obtain dental pulp stem cells; and S3, culturing the dental pulp stem cells by using a complete culture medium, when the confluence degree of the dental pulp stem cells reaches 60-80%, replacing the serum-free culture medium for continuous culture, collecting the dental pulp stem cell conditioned culture medium, and performing differential centrifugation to obtain the dental pulp stem cell exosome. According to the technical scheme, the regulating effect of the dental pulp stem cell exosome on endoplasmic reticulum stress and the mechanism of the dental pulp stem cell exosome for improving cartilage cell degeneration are explored.

Factor rich compositions produced from umbilical cord (UC) mesenchymal stem cells (MSCs) are described. Secretory UC MSCs in serum free culture conditions produce a factor rich conditioned medium which may be concentrated and filtered to obtain clinical grade products.

The application discloses immortalized chicken embryo epithelioid cells and a preparation method and application thereof, and the preparation method of the immortalized chicken embryo epithelioid cells comprises the following steps: extracting primary chicken embryo epithelial cells from a chicken embryo; and culturing the primary chicken embryo epithelioid cells in a conditional medium containing chicken Noggin protein for subculture, so as to obtain the immortalized chicken embryo epithelioid cells. The immortalized chicken embryo epithelioid cells prepared by the preparation method can be stably subcultured for a long time, can maintain stable proliferation ability after multiple subcultures, reduces individual differences, and improves the reliability and repeatability of experimental results. In addition, the preparation method does not introduce exogenous genes or change the genotype of the cells, ensures the stability at the gene level, and avoids the interference of the exogenous genes on the stability of the cell phenotype.

The subject invention pertains to compositions of a probe comprising two pH-reversible forms, including the boron dipyrromethene (BODIPY) fluorescent form and the non-fluorescent leuco-BODIPY form, for use in confocal imaging methods of intracellular vesicles as well as extracellular vesicles that are present in conditioned culture medium and / or secreted by living cells, without inducing severe cytotoxicity.

Tissue-specific progenitor cells representing the endoderm, ectoderm, and mesoderm lineages are generated from pluripotent sources. Progenitor cell generation is accomplished by contacting pluripotent stem cells with tissue-specific exosomes, microRNAs, proteins, and peptides, obtained by stressing the tissue under in vitro conditions. In vitro-generated tissue organoids are used as a source of "differentiation factors" for generating individualized progenitor cells. For example, to generate lung progenitor cells, allogeneic stem cells are implanted into decellularized cadaveric lung tissue to form lung structures, which are then exposed to various cellular stressors and conditioned media to differentiate the pluripotent stem cells into progenitor cells. Extracellular matrix can be obtained from decellularized structures seeded with regenerating cells and used to generate organoids, which are then subjected to stress to produce differentiation factors.

The invention belongs to the technical field of tissue engineering and aging resistance, and particularly relates to a composition for delaying cell aging and maintaining tissue homeostasis as well as a preparation method and application of the composition. Specifically, research finds that adipose tissues, skin tissues and the like are placed in a serum-free basal culture medium, adherent culture is performed, the obtained supernate is collected, and the obtained serum-free conditioned culture medium preparation can effectively inhibit aging of cells (such as ADSCs or HDFs) and maintain the functional steady state of the cells in in-vitro culture. Meanwhile, the culture medium product provided by the invention is also proved to be capable of inhibiting an IL-6 / STAT3 signal channel, and the channel is revealed to be a core mechanism of the anti-aging effect of the culture medium product, so that mechanism support is provided for the application of the culture medium product to cell aging resistance, and the culture medium product has good practical application value.

The utility model discloses a stem cell conditioned medium filter device which comprises a filter device shell, a filter tank is arranged in the filter device shell, a square sleeve is arranged on the lower side in the filter tank, a coarse-mesh filter screen is fixedly arranged in the square sleeve, and a filter screen is arranged in the coarse-mesh filter screen. A plurality of fine-hole filter screens fixed in the square sleeve are fixedly arranged below the coarse-hole filter screen; a sealing plate fixed in the square sleeve is arranged above the coarse-mesh filter screen, and a first stepping motor is fixedly mounted in the sealing plate. According to the utility model, the situation that the filter holes in the filter screen are blocked by cell debris, dead cells or metabolic wastes in the stem cell conditioned culture medium and even penetrate through the filter holes due to relatively intense impact generated between the filter screen and the stem cell conditioned culture medium when the filter screen is used for filtering the stem cell conditioned culture medium is prevented; through the technical scheme, the filtering effect of the equipment is improved.

The present application belongs to the technical field of stem cell exosome preparation. More specifically, it relates to adipose-derived stem cells (ADSCs), conditioned medium (CM) thereof, and preparation method and use thereof in cosmetics and drugs. The present application provides a mesenchymal stem cell exosome preparation for delaying aging, wherein the exosome preparation is a conditioned medium obtained by culturing mesenchymal stem cells in a low-osmotic induction combined with epi-pen stimulation environment. Tests show that the conditioned medium can significantly enhance the proliferation and migration ability of senescent HSFs cells, reduce the expression of cell beta-galactosidase, reduce the expression of cell-related aging genes p16 and p53, and reduce the generation of intracellular ROS, and has a significant anti-aging effect.

Disclosed herein is a pharmaceutical composition containing a neprilysin-overexpressing stem cell, a conditioned medium thereof, and an exosome isolated therefrom as an active ingredient for prevention or treatment of cognitive impairment. Having the functions of protecting nerve cells, inhibiting the accumulation of amyloid beta, known as a cause of cognitive impairment-related disease in animal models, increasing an expression level of acetyl choline, and improving cognitive functions in animal models, the neprilysin-overexpressing stem cells, the conditioned medium thereof, or the exosomes isolated therefrom according to the present disclosure can find advantageous applications in treating cognitive impairment.

To provide a method for generating secretomes using a serum-free medium.SOLUTION: A method for generating a secretome comprises: (a) culturing one or more progenitor cells in a first serum-free culture medium, the first serum-free culture medium comprising a basal medium, human serum albumin, and one or more growth factors; (b) removing the first serum-free culture medium from the one or more progenitor cells; (c) culturing the one or more progenitor cells in a second serum-free culture medium, the second serum-free culture medium comprising a basal medium but not comprising human serum albumin or growth factors; and (d) recovering the second serum-free culture medium after the culturing the step (c), thereby obtaining a conditioned medium comprising the secretome of one or more progenitor cells.SELECTED DRAWING: None

The invention discloses a preparation method of a bionic nanoparticle preparation of a antler stem cell conditioned culture medium, which comprises the following steps: S1, isolated culture of antler stem cells, preparation of P3 cells, and preparation of the antler stem cell conditioned culture medium ABPCs-CM from the P3 cells for later use; s2, synthesizing antler stem cell conditioned medium nanoparticles CM-NPS by adopting the antler stem cell conditioned medium ABPCs-CM obtained in the step S1; s3, preparing a neutrophile granulocyte membrane; s4, the antler stem cell conditioned medium nano-particles CM-NPS obtained in S2 and the neutrophile granulocyte membrane obtained in S3 are synthesized into the antler stem cell conditioned medium bionic nano-particle preparation, and the neutrophile granulocyte membrane bionic structure remarkably improves the targeting efficiency of the inflammation part.

The present invention provides compositions and methods for treating cardiac medical conditions such as heart failure caused by damaged myocardial tissue, as well as compositions and methods for addressing inflammatory and fibrotic processes observed in a wide range of medical conditions. [Solution] Embodiments of the present disclosure relate to compositions and methods of use relating to specific immortalized cells, including cardiomyocytes, obtained from pediatric or neonatal individuals. In specific embodiments, immortalized cells, or cell-derived conditioning media, or a partial or whole secretome thereof, are provided to individuals requiring it in an effective amount, either alone or in combination with cardiomyocytes.

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.

The present invention relates to a method for inducing, stimulating, and / or promoting the healing of a wound or damaged skin area, comprising topically treating a subject's skin surrounding the wound or damaged skin area with conditioned medium derived from a culture of umbilical cord mesenchymal stem cells. Similarly, the present invention relates to a method for preventing the formation and / or recurrence of wounds or damaged skin areas in a subject at risk of developing wounds or damaged skin areas, comprising topically treating the subject's skin with conditioned medium derived from a culture of umbilical cord mesenchymal stem cells. Furthermore, the present invention relates to a method for treating skin damage caused by rosacea, psoriasis, eczema, dermatitis, topical steroid withdrawal syndrome, epidermolysis bullosa, or fragile skin, comprising topically treating a subject's skin surrounding the wound or damaged skin area to be treated with conditioned medium derived from a culture of umbilical cord mesenchymal stem cells. The present invention is also directed to the conditioned medium, uses thereof, compositions comprising it, methods for producing the conditioned medium, and uses of these compositions.

The present disclosure provides methods related to tissue regeneration using human mesenchymal stem cell (MSC) / CD34+ hematopoietic stem / progenitor cell (HSPC) co-cultured total conditioned media. In particular, the present disclosure provides novel methods for treating and / or preventing damage to, trauma to, and / or loss of tissue.

The invention belongs to the field of biological medicine, and discloses a mesenchymal stem cell-derived exosome and active factor secretion composition and a directional preparation method thereof. According to the method, umbilical cord tissues of healthy mammals are used as raw materials, primary stem cells are separated through enzymolysis, and amplification and passage are conducted to 3-5 generations through an alpha-MEM culture medium; a serum-free induction culture medium containing multiple functional components is adopted for directional induction, a conditioned culture medium is collected, gradient centrifugation and ultrafiltration concentration are performed, and the protein concentration is calibrated to be 2 mg / mL, so that a finished product is obtained. The product can synergistically realize moisturizing, anti-wrinkle and tightening effects. Specifically, the exosome and active factor secreted composition is a composite system which is obtained by separating and concentrating mesenchymal stem cells from a conditioned medium of the mesenchymal stem cells after the mesenchymal stem cells are directionally induced by the method and is mainly composed of the exosome, growth factors self-secreted by the stem cells, cell factors and macromolecular active protein.

The method of hair transplant using dermal papilla and dermal sheath obtained from hair follicle as disclosed comprises the steps of: providing initial keratinocytes to be used for creating condition medium; creating and collecting condition medium from said keratinocytes; collecting hair follicle cells containing dermal papilla and dermal sheath from patients; culturing said cells as obtained from patients using condition medium collected from the prior step; injecting cultured cells in the predetermined amount and the predetermined ratio of dermal papilla cells and dermal sheath cells in the predetermined amount area characterized in that the step of injecting and collecting condition medium from said keratinocytes comprises the step of applying a solution obtained from the mixture of defined keratinocyte-serum free medium (DK-SFM) and Mesenchymal stem cell culture medium (MSC) to said initial keratinocytes, removing defined keratinocyte-serum free medium (DK-SFM), and extracting a condition medium from the said mixture and wherein the step of culturing hair follicle cells as obtained from patients comprises the step of applying condition medium extracted and MSC in the ratio of 1:1.

The invention relates to a method for evaluating the biological value of mesenchymal stem cells, and belongs to the technical field of biology. The method comprises the following steps: firstly, respectively collecting conditioned culture media of mesenchymal stem cells from different donor sources and mesenchymal stem cells expressed by silence TSP2, and detecting the concentration of TSP2 in the culture media by adopting an enzyme-linked immunosorbent assay method; furthermore, inflammatory factors are used for inducing cartilage cells, a conditioned culture medium is replaced for culture for 24 h, then the cells are collected, messenger RNA (mRNA) expression change of II-type collagen of the cells is detected, and the correlation between the TSP2 concentration of the conditioned culture medium and the mRNA expression abundance of the II-type collagen of the cartilage cells is established; the TSP2 concentration in the mesenchymal stem cell culture supernatant is measured, the TSP2 concentration can be used for evaluating the biological value of the mesenchymal stem cells in cartilage regeneration and protection in treatment of osteoarthritis and other degenerative diseases, and a reliable basis is provided for standardization and quality control of mesenchymal stem cell treatment.

The invention relates to the technical field of pharmaceutical compositions, in particular to an atomization composition and application thereof. An aerosolized composition comprising: A) an MSC-derived active ingredient selected from any one or a combination of (i) MSC exosomes / extracellular vesicles, (ii) an MSC conditioned culture solution concentrate, (iii) an MSC lysate, (iv) an MSC-derived soluble factor, (v) an immobilized / embedded protected MSC or fragment thereof; b) ergothioneine (EGT) or a pharmaceutically acceptable salt thereof; and C) a pharmaceutically acceptable carrier / buffer system making it suitable for aerosol inhalation administration. The exosome is combined with ergothioneine, and the effectiveness of sleep improvement is remarkably improved through mechanism synergy of nerve regulation and cell protection; the introduction of the atomization drug delivery technology breaks through the limitation of the traditional therapy from multiple dimensions of non-invasiveness, accurate delivery, individualized treatment, accessibility and the like, so that wider crowds (including special crowds) can safely, conveniently and efficiently receive treatment.

A technology regarding the production, formulation and use of conditioned media and the factors included therein is disclosed. The conditioned media may be inoculated with animal cells, plant cells and any combination thereof. The inoculations may occur simultaneous or at different times. Cells retrieved from different areas of the animal and / or the plant may also be cultured together to form conditioned media and associated growth factors.

The present disclosure is generally directed to the use of biomaterial scaffolds engineered with SA-FasL for the transplantation of stem cell derived β-cells as a treatment for Type I diabetes. Early engraftment post-transplantation and subsequent maturation of these β-cells may be limited by the initial inflammatory response, which impacts the ability to sustain normoglycemia at long times. The survival and development of immature hPSC-derived β-cells transplanted on poly(lactide-co-glycolide) (PLG) microporous scaffolds into the peritoneal fat, a site being considered for clinical translation, was investigated. The scaffolds were modified with biotin for binding of a streptavidin-FasL (SAFasL) chimeric protein to modulate the local inflammatory microenvironment. The presence of FasL impacted infiltration of monocytes and neutrophils and altered their phenotypic response. Conditioned media generated from scaffolds explanted at day 4 did not impact hPSC-derived β-cell survival and maturation in vitro, which was not observed with unmodified scaffolds. Following transplantation, β-cell viability and differentiation were improved with SA-FasL modification. A sustained increase in insulin positive cell ratio was observed with SA-FasL modified relative to unmodified scaffolds. These results demonstrate that SA-FasL-modified scaffolds can mitigate initial inflammatory response and enhance β-cell engraftment and differentiation.