60 results about "Pre adipocytes" patented technology

The present invention relates generally to a tissue preparation including tissue cells and extracts thereof useful for promoting or facilitating the growth, development and differentiation of cells and tissues. More particularly, the present invention provides muscle-derived material comprising intact or extracted extracellular matrix and / or cells as well as cytokines, growth factors and other components. The muscle preparations of the present invention resemble basement membrane and are derived from cellular-based material. The muscle preparation may be used in vitro or in vivo as inter alia, a cellular scaffold in various tissue engineering applications and in other cell culture systems for nurturing and enriching a range of cell types including, but not limited to, precursor and stem cells such as pre-adipogenic cells. The muscle preparation is also useful as a base for creams, such as in the cosmetic and topical therapeutic industries and as a matrix or additive in the food industry.

Methods for inducing or accelerating a healing process of a damaged skin or skin wounds are described. The methods include administering to the skin cells colonizing the damaged skin or skin wound a therapeutically effective amount of an adipokine, an adipocyte or preadipocyte modulator, adipocytes, preadipocytes, or stem cells, or transforming the skin cells colonizing the damaged skin or skin wound such as to express and secrete an adipokine, thereby inducing or accelerating the healing process of the damaged skin or skin wound.

The present invention provides methods and compositions for the consistent and quantitative differentiation of human preadipocytes isolated from adipose tissue into adipocytes bearing biochemical, genetic, and physiological characteristics similar to that observed in isolated primary adipocytes. The methods of the invention comprise incubating isolated human preadipocytes, plated at least about 25,000 cells / cm2, in a medium containing, glucose, a cyclic AMP inducer such as isobutylmethylxanthine or forskolin, a glucocorticoid or glucocorticoid analogue, insulin or an insulin analogue and a PPARγ agonist or a RXR agonist. The compositions of the invention include media for the differentiation of human preadipocytes, human adipocytes differentiated by the methods of the invention and transfected adipocytes.The present invention also provides methods for determining the ability of a compound to affect the differentiation of human preadipocytes to adipocytes, for determining the ability of a compound to act as a PPARγ antagonist. a glucocorticoid, a glucocoticoid analogue, or an insulin analogue, for transfecting cultured human adipocytes, and as a means to identify novel polypeptides secreted from human adipocytes into the conditioned medium. The methods and compositions have use in the drug discovery of compounds having relevance to the disease states of diabetes, obesity, and cardiovascular disease and in the studies of these diseases.

The present invention is directed to methods and agents for modulating the differentiation potential and / or proliferation of preadipocytes. More particularly, the present invention discloses methods and agents for modulating a fibroblast growth factor (FGF) signaling pathway, especially the FGF-1 or FGF-2 signaling pathway, for treating or preventing adiposity-related conditions including, but not limited to, obesity, lipoma, lipomatosis, cachexia or lipodystrophy or the loss of adipose tissue in trauma or atrophic conditions.

The present invention provides methods and compositions for the consistent and quantitative differentiation of human preadipocytes isolated from adipose tissue into adipocytes bearing biochemical, genetic, and physiological characteristics similar to that observed in isolated primary adipocytes. The methods of the invention comprise incubating isolated human preadipocytes, plated at least about 25,000 cells / cm2, in a medium containing, glucose, a cyclic AMP inducer such as isobutylmethylxanthine or forskolin, a glucocorticoid or glucocorticoid analogue, insulin or an insulin analogue and a PPARγ agonist or a RXR agonist. The compositions of the invention include media for the differentiation of human preadipocytes, human adipocytes differentiated by the methods of the invention and transfected adipocytes. The present invention also provides methods for determining the ability of a compound to affect the differentiation of human preadipocytes to adipocytes, for determining the ability of a compound to act as a PPARγ antagonist. a glucocorticoid, a glucocoticoid analogue, or an insulin analogue, for transfecting cultured human adipocytes, and as a means to identify novel polypeptides secreted from human adipocytes into the conditioned medium. The methods and compositions have use in the drug discovery of compounds having relevance to the disease states of diabetes, obesity, and cardiovascular disease and in the studies of these diseases.

The present invention relates to preadipocyte strains that maintain replicative potential and adipogenic capacity. In particular, the invention relates to preadipocytes engineered to express telomerase reverse transcriptase (TERT). Use of the cells as research tools, in screening assays, and as therapeutic and / or clinical reagents is also described.

The subject invention pertains to supercritical carbon dioxide extracts of Commiphora mukul resin (guggul), which can be modified or not modified by some ethanol addition; methods for their production; and methods of use, such as inhibiting HMG-CoA reductase, inhibiting transformation of pre-adipocytes to adipocytes, inhibiting triglyceride storage, promoting insulin sensitivity in adipocytes, treatment of disorders (for example, hypercholesterolemia, hyperlipidemia, hyperglycemia, obesity, metabolic syndrome, cardiovascular disease, atherosclerotic heart disease, autoimmune disorder, insulin resistance, leptin resistance, arthritis, cell proliferation disorder, such as cancer and atherosclerosis; damaged skin, sores, cuts, rashes, bruises, dryness, burns, sunburn, radiation burn, and infection), and regulating or suppressing appetite.

The present invention provides methods and compositions for the induction of expression of UCP1 independent of lipid accumulation. The invention, in particular, features methods for converting FGF receptive cells, e.g., preadipocyte cells, into energy consuming cells through FGF-mediated UCP1 expression. The invention further provides methods and compositions for treating metabolic disorders with an FGF receptor agonist, (e.g., an FGF protein, or fragment thereof, a nucleic acid encoding an FGF protein, an FGF mimetic, an anti-FGF receptor agonist antibody, or antigen binding fragment thereof), or a cell contacted with an FGF receptor agonist, including FGF6.

The present invention describes method for culturing preadipocytes isolated ex vivo, the method including introducing preadipocytes into a three dimensional support matrix, and allowing the cells to differentiate in vitro into adipocytes within the support matrix. The matrix may be a collagen matrix. The method may be used for investigating the development of stem cells, or for investigating the response of adipocytes to stimuli. The method provides a system whereby adipocytes with biological properties resembling those in vivo can be grown in vitro.

The invention discloses a method for culturing and inducing tilapia mossambica peritoneal preadipocytes. The method comprises the following steps: a) resuspending cells by using a DMEM / F12 culture medium with 10%-15% of fetal calf serum, inoculating the cells into a cell plate, and culturing for 7 days at the temperature of 20-30 DEG C under the condition of 5% CO2; and b) replacing an induction culture medium for sequentially culturing, after inducing for 7-14 days, carrying out oil red O staining of the cells, wherein the fuzzy cell membrane edges can be seen under the microscope, and the interiors of the cells are stained to be red lipid droplets. The invention also discloses a passage method of tilapia mossambica peritoneal preadipocytes. A proliferation culture medium is used for culturing the preadipocytes for 14-20 days, then the preadipocytes are inoculated according to 2 *10<5> to 3*10<5> per milliliter, and the cells can overgrow on the cell plate after the cells are cultured for 2-3 days. The method is capable of successfully carrying out in-vitro culture, induction and passage of the tilapia mossambica peritoneal preadipocytes; the technical reference is provided for the research of the tilapia mossambica lipid metabolism.

The invention provides methods and compositions relating to molecular targets identified as being capable of increasing or decreasing thermogenic potential in cells, including preadipocytes. Included in the invention are methods and compositions relating to inhibiting or suppressing the activity of an uncoupling protein 1 (UCP1) negative regulator, such as cardiac actin 1 (ACTC1), somatostatin receptor 1 (SSTR1), FAT atypical cadherin 1 (FAT1), and protein tyrosine phosphatase receptor type B (PT-PRB). Also included in the invention are methods and compositions relating to activating a UCP1 positive regulator, such as phosphatidylinositol-3,4,5-triphosphate-dependent Rac exchange factor 1 (PREX1), cortactin binding protein 2 (CTTNBP2), doublesex and mab-3-related transcription factor-like family A1 (DMRTA1), and endothelin receptor type B (ENDRB). The invention also provides methods and compositions relating to enrichment of cells having thermogenic potential based on cell surface markers, e.g., CD29, identified as being predictive of such.

The invention provides wine-steamed Chinese goldthread as well as a preparation method and application thereof. The wine-steamed Chinese goldthread as an insulin sensitizer and a PPAR gamma activator has the functions of inhibiting the differentiation of 3T3-L1 pre-adipocytes, improving the utilization rate of glucose of the 3T3-L1 pre-adipocytes, reducing blood fat, improving insulin resistance and treating diabetes mellitus. The wine-steamed Chinese goldthread and extracts thereof have the functions of inhibiting the differentiation of the 3T3-L1 pre-adipocytes, improving the utilization rate of the glucose of the 3T3-L1 pre-adipocytes and reducing the blood fat and the application in improving insulin resistance or treating diabetes mellitus, have a better specific effect than that of berberine hydrochloride and crude Chinese goldthread as well as extracts thereof, and the safety obviously higher than that of the crude Chinese goldthread.

Using electroporation, it is possible to activate the natural reprogramming potential of living Xenopus laevis oocytes and pass it on to donor cells placed with eggs in one electroporation chamber. We demonstrated that co-electroporation at 150 v / cm / 25 μF of mature oocytes with ˜105 cells / ml of suspension of various normal and cancerous human cell lines, such as bone marrow stromal cells, foreskin fibroblasts, pre-adipocytes, CD4+ T-lymphocytes, cheek cells, cervical carcinoma (HeLa) cells and breast adenocarcinoma (MCF-7) cells, reprograms donor cells into iPSc-like cells, which form colonies on irradiated MEF feeders. The iPSc-like cells generated by this study resemble human embryonic stem cells in colony morphology and expression of stem cell-associated transcription factors, including Oct3 / 4, Nanog, SOX-2, Rex-1, TRA-1-60 and SSEA-1. New method obviates the use of retroviral or lentiviral gene delivery vectors and other “non-parental” reprogramming approaches.

The invention features methods of obtaining high-yield, essentially pure human predipocyte cultures. Cultures obtained according to the instant methodology are also featured as are methods of identifying adipogenic modulatory agents, e.g., high-throughput screening assays.

The present invention provides compositions comprising an effective amount of xanthohumol and honokiol. The present invention comprises compositions comprising an effective amount of xanthohumol and genistein. The present invention comprises compositions comprising an effective amount of xanthohumol and guggulsterone. The compositions of the present invention may be effective to decrease mature adipocytes viability, induce apoptosis of mature adipocytes, increase lipolysis in mature adipocytes, and / or reduce adipogenesis during the maturation of pre-adipocytes. The present invention further provides methods of treating obesity, diabetes, osteoporosis or bone disorders in a subject, which comprise administering to the subject compositions comprising an effective amount of xanthohumol and honokiol, guggulsterone, or genistein.

A method for the identification of a composition useful in the treatment of an overweight or obese person to reduce the person's body mass index is provided which comprises obtaining an extract of an ethnobotanical plant, and evaluating the activity of the extract in an assay selected from the group consisting of a lipolysis assay, an assay that measures the amount of glycerol introduced by a cell into a suspension medium of the cell, an adipocyte differentiation assay, an assay that measures the level of the enzyme glycerol-3-phosphate dehydrogenase, an assay that measures the inhibition of differentiation of preadipocytes to adipocytes, an assay that measures the accumulation of lipid in an adipocyte, an assay that measures the de-differentiation of adipocytes into preadipocytes, and combinations thereof. Sclareol and sclareol-like compounds and sclareolide and sclareolide-like compounds are identified as useful in the treatment of overweight and obese conditions and disorders and conditions associated with adipose tissue abnormalities. These compounds can be formulated for oral administration.

Disclosed is a method of identifying genes that are over-expressed in adipose tissue as compared to pre-adipocyte tissue or other tissues comprising performing differential gene expression analysis between the white adipose tissue (WAT) or stromal vascular tissue (SVT) from any two different mice selected from the group consisting of wild-type, HMGI-C − / −, ob / ob, and HMGI-C − / − ob / ob genotype mice. Based on this method a number of nucleotide sequences are identified whose expression is adipocyte specific. The identified nucleotide sequences and their corresponding polypeptides are then used to prevent adipogenesis, to treat diabetes, and to screen for small-molecules that can modulate or prevent adipogenesis and to treat diabetes.

Provided are live, artificial, skin substitute products and methods of making and using the same, such as for wound treatment and compound testing, including compound testing for efficacy, toxicity, penetration, irritation and / or metabolism testing of drug candidates or compositions such as cosmetics. Described herein is an artificial mammalian skin substitute product, comprising: (a) optionally, but in some embodiments preferably, a first (“hypodermis-like”) layer comprising live mammalian adipocytes (e.g., induced pre-adipocytes) in a first hydrogel carrier; (b) a second (“dermis-like”) layer contacting or directly contacting the first layer and comprising live mammalian fibroblast cells and' live mammalian follicle dermal papilla cells in combination in a second hydrogel carrier; (c) a third (“epidermis-like”) layer contacting or directly contacting the second layer (i.e., on the opposite side thereof as the first layer, so that the second layer is sandwiched between the first and third layers when the first layer is present), the third layer comprising live mammalian keratinocytes and live mammalian melanocytes in combination in a third hydrogel carrier.

The present invention provides a method for the determination of leptin in a sample, based on the detection of leptin-induced angiopoietin in certain cells or cell lines, such as Swiss 3T3 F442A mouse preadipocytes or human bone marrow stromal cell line hMS2-12 -2 Detection of expression. The detection of leptin is as follows: cells are transfected with a vector containing a reporter gene (such as green fluorescent protein (GFP)) under the control of angiopoietin-2 promoter.

The invention discloses a transcription factor C / EBPZ for regulating adipocyte formation and application thereof, belongs to the field of animal molecular genetics and developmental biology, and discloses application of the transcription factor C / EBPZ for regulating and controlling chicken adipocyte formation. The invention discloses an expression mode of chicken C / EBPZ in various tissues and an expression rule of C / EBPZ in a chicken adipose tissue growth and development process by utilizing an RT-PCR technology. According to the invention, an overexpression vector of the transcription factoris constructed, and the C / EBPZ transcription factor is verified to regulate and control the differentiation and proliferation of chicken preadipocytes; the invention further discloses an action mechanism of the transcription factor C / EBPZ in chicken adipose tissue formation. The invention provides a novel application of a C / EBPZ transcription factor in regulation and control of chicken adipocyte formation, and the C / EBPZ transcription factor has practical value in the fields of chicken genetic breeding and animal nutrition.

The subject invention pertains to supercritical carbon dioxide extracts of Commiphora mukul resin (guggul), which can be modified or not modified by some ethanol addition; methods for their production; and methods of use, such as inhibiting HMG-CoA reductase, inhibiting transformation of pre-adipocytes to adipocytes, inhibiting triglyceride storage, promoting insulin sensitivity in adipocytes, treatment of disorders (for example, hypercholesterolemia, hyperlipidemia, hyperglycemia, obesity, metabolic syndrome, cardiovascular disease, atherosclerotic heart disease, autoimmune disorder, insulin resistance, leptin resistance, arthritis, cell proliferation disorder, such as cancer and atherosclerosis; damaged skin, sores, cuts, rashes, bruises, dryness, burns, sunburn, radiation burn, and infection), and regulating or suppressing appetite.

The invention provides application of golden thread in preparing medicaments, such as an insulin sensitizer and a PPAR-gamma agonist, in particular relates to application in restraining 3T3-L1 pre-adipocyte cell differentiation, enhancing the utilization ratio of 3T3-L1 pre-adipocyte glucose or reducing blood fat and improving insulin resistance or diabetes medicaments. The invention also provides a medicinal composition for restraining 3T3-L1 pre-adipocyte cell differentiation and enhancing the utilization ratio of glucose. The golden thread and the extract thereof have the advantages of restraining 3T3-L1 pre-adipocyte cell differentiation, improving insulin resistance and treating diabetes and complicating diseases thereof, and achieving remarkable medicament effect which is superior to that of berberine hydrochloride and crude golden thread.

The invention discloses a C / EBPZ gene promoter and an application thereof, and belongs to the field of gene engineering and molecular biology. The C / EBPZ gene promoter is any one of A1-A6 sequences of which the nucleotide sequences are as shown in SEQ ID NO: 1-SEQ ID NO: 6. Six chicken C / EBPZ gene promoters are obtained by using DNA extracted from chicken whole blood as a template and using primers for PCR amplification and restriction enzyme digestion, and tests prove that the six promoters all have promoter activity in chicken preadipocytes and are regulated by a specific transcription factor KLF2, so that the promoter has potential application value in revealing a transcription regulation mechanism of chicken C / EBPZ, breeding low-abdominal-fat broilers, revealing molecular mechanisms of various human diseases and developing drugs for targeting C / EBPZ expression.

Provided are live, artificial, skin substitute products and methods of making and using the same, such as for wound treatment and compound testing, including compound testing for efficacy, toxicity, penetration, irritation and / or metabolism testing of drug candidates or compositions such as cosmetics. Described herein is an artificial mammalian skin substitute product, comprising: (a) optionally, but in some embodiments preferably, a first (“hypodermis-like”) layer comprising live mammalian adipocytes (e.g., induced pre-adipocytes) in a first hydrogel carrier; (b) a second (“dermis-like”) layer contacting or directly contacting the first layer and comprising live mammalian fibroblast cells and' live mammalian follicle dermal papilla cells in combination in a second hydrogel carrier; (c) a third (“epidermis-like”) layer contacting or directly contacting the second layer (i.e., on the opposite side thereof as the first layer, so that the second layer is sandwiched between the first and third layers when the first layer is present), the third layer comprising live mammalian keratinocytes and live mammalian melanocytes in combination in a third hydrogel carrier.

The invention provides application of wine-roasted Chinese goldthread in preparing medicaments containing insulin sensitizers and PPAR gamma activators. The wine-roasted Chinese goldthread has the functions of inhibiting the differentiation of 3T3-L1 pre-adipocytes, improving the utilization rate of glucose of the 3T3-L1 pre-adipocytes or reducing blood fat as the insulin sensitizers and the application in improving insulin resistance or treating diabetes mellitus. The invention also provides a medicine composition having the functions of inhibiting the differentiation of 3T3-L1 pre-adipocytes and improving the utilization rate of the glucose of the 3T3-L1 pre-adipocytes. The wine-roasted Chinese goldthread and extracts thereof are the medicaments having the function of inhibiting the differentiation of 3T3-L1 pre-adipocytes, can be used for improving insulin resistance and treating the diabetes mellitus and complications thereof and have a specific effect better than that of berberine hydrochloride and crude Chinese goldthread.

The present invention is directed to constructs, compositions and methods for modulating platelet-type 12 lipoxygenase (12-LO) in adipose tissue in vivo. Specifically, the invention provides constructs encoding expression-inhibiting oligonucleic acids, e.g., antisense and RNA interfering (RNAi) molecules, targeted to a platelet-type 12-LO gene or a transcript thereof, which are capable of reducing or silencing platelet-type 12-LO expression specifically in adipocytes and pre-adipocytes. Vectors of these constructs (including, but not limited to, viral vectors), compositions of them and methods of using same for the treatment and amelioration of conditions associated with excess fat cell mass and obesity are also provided.

The invention discloses application of a chicken CTGF gene in inhibition of chicken preadipocyte differentiation. The nucleotide sequence of the chicken CTGF gene is as shown in SEQ ID NO.1, and the coded amino acid sequence of the chicken CTGF gene is as shown in SEQ ID NO.2. According to the invention, through a method for adding an exogenous CTGF recombinant protein and constructing an endogenous pCMV-CTGF overexpression vector, an oil red O technology, a Real-time PCR technology and a Western blot technology are utilized to prove that the CTGF gene can inhibit chicken preadipocytes from being differentiated into mature adipocytes from the lipid droplet deposition capability, the transcriptome level and the proteome level of the chicken adipocytes, and thus, the invention provides novel application of the CTGF gene in regulation and control of the chicken preadipocyte differentiation process, and the CTGF gene has practical value in the fields of chicken genetic breeding and animal nutrition.

The invention provides novel polynucleotides and polypeptides encoded by such polynucleotides and mutants or variants thereof that correspond to a novel human secreted Pref-1-like polypeptide. These polynucleotides comprise nucleic acid sequences isolated from cDNA library from adult human brain (Hyseq clone identification numbers 7958079 (SEQ ID NO:1)). Other aspects of the invention include vectors containing processes for producing novel human secreted Pref-1-like polypeptides, and antibodies specific for such polypeptides.

The present invention provides compositions comprising an effective amount of xanthohumol and honokiol. The present invention comprises compositions comprising an effective amount of xanthohumol and genistein. The present invention comprises compositions comprising an effective amount of xanthohumol and guggulsterone. The compositions of the present invention may be effective to decrease mature adipocytes viability, induce apoptosis of mature adipocytes, increase lipolysis in mature adipocytes, and / or reduce adipogenesis during the maturation of pre-adipocytes. The present invention further provides methods of treating obesity, diabetes, osteoporosis or bone disorders in a subject, which comprise administering to the subject compositions comprising an effective amount of xanthohumol and honokiol, guggulsterone, or genistein.

A method for identifying an agonist exhibiting molecular or pharmacologic inhibition which is effective against the activity of at least one of iPLA2β and iPLA2γ which comprises culturing 3T3-L1 cells and transfecting them with negative control siRNA, siRNA directed against iPLA2β or siring directed against iPLA2γ prior to induction or during to differentiation or pharmacologic inhibition and observing for whether that down regulation of iPLA2β or iPLA2γ inhibits adipocyte differentiation.