45 results about "Rat Bone Marrow" patented technology

The invention discloses a magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2 and a preparation method thereof. The method for preparing the magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2 comprises the steps of: firstly preparing hydrophobic monodisperse Fe3O4 nano-particles by adopting a chemical oil-phase high-temperature method, and modifying the surfaces of the hydrophobic Fe3O4 nano-particles to ensure that the hydrophobic Fe3O4 nano-particles are dispersed in a water phase; preparing luminescent CdTe quantum dots of which the surfaces are provided with carboxyl groups, and precipitating the luminescent CdTe quantum dots on the surfaces of the magnetic Fe3O4 nano-particles through the co-precipitation; then utilizing ligand exchange to modify a silane coupling agent on the surfaces of the luminescent CdTe quantum dots; and finally forming an outermost SiO2 coating layer through silane or silicon ester hydrolysis. The diameter of the magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2 is between 30 and 50nm; the magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2has double functions of magnetism and fluorescence at the same time, has strong and durable fluorescence intensity after labeling rat bone marrow-derived mesenchymal stem cells, and apparently reduces cellular magnetic resonance signals. The particle has broad application prospect in the fields such as biological labeling, bioseparation and the like.

Intravenous administration of bone marrow cells collected from rat bone marrow to a rat cerebral infarction model was found to be effective in treating cerebral infarction. Human and murine bone marrow stem cells showed similar effects. Mesenchymal cells such as bone marrow cells, cord blood cells, or peripheral blood cells can be used as agents for in vivo administration against cranial nerve diseases.

Intravenous administration of bone marrow cells collected from rat bone marrow or peripheral blood to a rat cerebral infarction model was found to be effective in treating cerebral infarction. Human and murine bone marrow stem cells showed similar effects. Mesenchymal cells such as bone marrow cells, cord blood cells, or peripheral blood cells can be used as agents for in vivo administration against cranial nerve diseases.

The invention discloses a recombinant adenovirus carrying a rat retinoic acid receptor (RAR) gamma gene. The gene of the recombinant adenovirus is derived from a rat mesenchymal stem cell and has an SEQID1 sequence. A construction method comprises the following steps: acquiring a target gene RAR gamma; constructing and identifying a recombinant adenovirus plasmid pAd-RAR gamma; and carrying out package, amplification, purification and titer detection on the recombinant adenovirus pAd-RAR gamma in a human embryonic kidney (HEK) 293 cell. In the invention, the rat mesenchymal stem cell can be efficiently infected and the RAR gamma gene can be efficiently expressed, thereby well solving the problem that a general eukaryotic vector is difficultly led in the RAR gamma cell through transfection of liposome; the physical and chemical properties of the used adenovirus vector are stable, thus preparation and operation are easy; the transfection efficiency of the gene is high, and exogenous gene expression level is high; and the obtained adenovirus is replication-defective, is not combined in a gene group of a host cell, has a high biological safety, and is widely suitable for various in vitro cells and in vivo experiment studies.

The invention belongs to the technical field of traditional Chinese medicines, discloses a drug for inducing bone marrow mesenchymal stem cell (MSC) differentiation to cardiomyocyte and a detection method thereof. The method includes: preparation of drug serum of Zhejiang radix ophiopogonis; preparation of cardiomyocyte extraction solution and lysate; separation and culture of rat MSCs; identification of surface markers of rat MSCs by a flow cytometer; culture of rat MSCs; detection of the expression of cardiomyocyte-associated proteins CTnI and CX43 by immunofluorescence; and detection of theexpression of NKx2.5, GATA-4 and beta-MHC mRNA by a qPCR method. The method disclosed by the invention establishes a high-efficiency and systematic research mode for batch searching and research of traditional Chinese medicine with potential inducibility, lays a foundation for perfecting the screening and development system of traditional Chinese medicine inducer, provides a new idea for the research of traditional Chinese medicine induction, has great significance for improving the success rate of autologous transplantation of MSCs, reducing the mortality of patients, and preventing the occurrence and development of diseases such as coronary heart failure.

Cellulose and sulfated cellulose fibrous meshes exhibiting robust structural and mechanical integrity in water were fabricated using a combination of electrospinning, thermal-mechanical annealing and chemical modifications. The sulfated fibrous mesh exhibited higher retention capacity for human recombinant bone morphogenetic protein-2 than the cellulose mesh, and the retained proteins remained biologically active for at least 7 days. The sulfated fibrous mesh also more readily supported the attachment and osteogenic differentiation of rat bone marrow stromal cells in the absence of osteogenic growth factors. These properties combined make the sulfated cellulose fibrous mesh a promising bone tissue engineering scaffold.

The invention discloses a recoverable immortalized rat bone marrow mesenchyme stem cell. The cell carries an SV40T (Simian Virus 40T) gene and a hygromycin resistance gene and two ends of the SV40T gene also have orthokinetic Loxp sites. The preparation method of the recoverable immortalized rat bone marrow mesenchyme stem cell comprises the following steps of: co-transfecting an HEK293 (Human Embryo Kidney 293) cell by SSR69 (Simple Sequence Repeat 69) plasmids and pAmhpo plasmids and screening by the hygromycin to obtain the recoverable immortalized rat bone marrow mesenchyme stem cell with hygromycin resistance; the cell has the same biological property with a primary rat and also has the advantages of strong external multiplication capacity, stable biological property, capability of being recovered to a previous state, good biological safety and the like; and the cell provided by the invention can be used as a seed cell induced and differentiated by nerves and a seed cell for treating hypoxicischemic brain damage of a newly-born rat.

Cellulose and sulfated cellulose fibrous meshes exhibiting robust structural and mechanical integrity in water were fabricated using a combination of electrospinning, thermal-mechanical annealing and chemical modifications. The sulfated fibrous mesh exhibited higher retention capacity for human recombinant bone morphogenetic protein-2 than the cellulose mesh, and the retained proteins remained biologically active for at least 7 days. The sulfated fibrous mesh also more readily supported the attachment and osteogenic differentiation of rat bone marrow stromal cells in the absence of osteogenic growth factors. These properties combined make the sulfated cellulose fibrous mesh a promising bone tissue engineering scaffold.

The invention relates to a RNA interference carrier used for osteoporosis and mesenchymal stem cells. The RNA interference carrier is constructed by introducing a DNA fragment which uses peroxisome proliferator-activated receptor Gamma (PPAR Gamma) as an action target and can express short chain RNA (shRNA) so as to build a slow virus expression carrier in a framework plasmid expressed based on the short chain RNA of slow virus; the constructed RNA interference carrier can express a special short chain RNA aiming at the PPAR Gamma, thereby reducing the expression of the PPAR Gamma. The invention also provides rat mesenchymal stem cells transfected by the RNA interference carrier. The invention not only provides a new way for preparing the medicine for treating the osteoporosis, but also provides a better experiment platform for the development of other stem cell-origin cell therapy and tissue engineering products.

Intravenous administration of bone marrow cells collected from rat bone marrow or peripheral blood to a rat cerebral infarction model was found to be effective in treating cerebral infarction. Human and murine bone marrow stem cells showed similar effects. Mesenchymal cells such as bone marrow cells, cord blood cells, or peripheral blood cells can be used as agents for in vivo administration against cranial nerve diseases.

The invention belongs to the medical field of biological tissue engineering, and particularly relates to a sulfonated polyetheretherketone-based bone repair composite material. The sulfonated polyetheretherketone-based bone repair composite material takes sulfonated polyetheretherketone as a carrier, and nano protein is loaded on the surface of the sulfonated polyetheretherketone-based bone repair composite material. The preparation method comprises the following steps: sulfonating polyether-ether-ketone with an acid solution, washing, carrying out water thermalization, and drying to obtain sulfonated polyether-ether-ketone; and dropwise adding the nano protein solution to the surface of the sulfonated polyether-ether-ketone, freeze-drying, freeze-drying, and disinfecting to obtain the sulfonated polyether-ether-ketone-based bone repair composite material. The product provided by the invention has good biocompatibility, can promote adhesion, proliferation and differentiation of SD rat bone marrow mesenchymal stem cells through test verification, also has excellent properties of oxidative stress resistance, antibacterial property and the like, and has the advantages of easily available raw materials, simple synthesis and preparation, simple and convenient process, high preparation efficiency and realization of large-scale production.

The invention discloses a method of induced differentiation of rat bone marrow mesenchymal stem cells into adipocytes, which includes: choosing and inoculating third-generation bone marrow mesenchymal stem cells into a cell culture flask according to a concentration of 1 multiplied by 106mL<-1>, and after the cells attaching to a wall grow until cell density reaches 80 percent, adding different concentrations of oleic acid as inducers; according to a volume proportion of 10 percent, adding fetal bovine serum into a basic culture medium DMEM (Dulbecco modified Eagle medium), and respectively adding 25mM of oleic acid, 50mM of oleic acid, 100mM of oleic acid and 200mM of oleic acid as induction culture media for forming adipocytes; replacing the solution once every three days, and with a cell culture flask without a induction culture solution as a control, performing oil red O staining assay after 3 weeks. According to the method, different concentrations of oleic acid can be used to directly induce adipocytes, a novel induction method is established, the configuration of the method is simpler, the induction speed is high, and the induction rate is high. In addition, the method can be used for inducing adipocytes of different sizes and researching the functional differences of the adipocytes of different sizes.

The invention provides a immortal stromal cell systme of rat's marrow and preparation thereof. Immortal stromal cell of rat's marrow can be obtained in short time by the inventive method. The inventive hTERT immortal stromal cell systme of rat's marrow is stable, can be sequentially passed through generations and keep diploid karyotype and characteristics of normal stromal cell of rat's marrow, has extremely strong proliferation ability, and realize immortalization. Immortal stromal cell system of rat's marrow established by the inventive method can conveniently and quickly obtain large amount of stromal cell of rat's marrow with diploid karyotype. This no doubt has great promotion to research of introduction and bone marrow stromal cell in vitro and in vivo.

The invention discloses a method for inducing gelation and biomimetic mineralization of a fibroin solution by alkaline phosphatase. According to the invention, the micromolecular polypeptide which issensitive to ALP and has good biocompatibility and self-assembly property is introduced as a gelator precursor; phosphoric acid groups on molecules can be removed under the catalytic action of ALP togenerate NY, supermolecule self-assembly is triggered, and therefore SF co-self-assembly is synergistically induced, and finally SF hydrogel is rapidly formed. ALP wrapped in an SF-NY hydrogel networkstill maintains the catalytic activity of the SF-NY hydrogel network, and free phosphate ions are released by catalyzing beta-glycerophosphate so that formation of apatite minerals is induced in thegel. Due to the mild gelation process and the formation of apatite minerals in the gel matrix, the biomimetic mineralized SF gel can be used as a biomimetic scaffold to promote the adhesion, proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro, and can also promote the natural healing of femoral defects in a rat model.

The invention relates to a method for extracting mesenchymal stem cells. The method includes the steps: performing pretreatment to obtain a thigh bone and a shin bone; flushing the thigh bone and theshin bone, extracting marrow cavity contents to obtain flushing liquid containing crumby marrow cavity contents; and collecting stem cells in the flushing liquid; blowing and beating remaining crumbymarrow cavity contents to obtain single-cell suspensions, and collecting stem cells in the single-cell suspensions; grafting and culturing the acquired stem cells. The method can be used for extracting mesenchymal stem cells of rats, and the acquired mesenchymal stem cells are applied to research of cell biology. The method has the advantages of simplicity in operation, low cost, pollution probability and stem cell loss, more cell extraction amount, high purity, good repeatability and the like, and the method is of great significance for biological research in the field.

The present invention provides an application of Rspo1 to induce differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells. Adenoviruses are used as a vector, Rspo1 is transfected into the rat bone marrow mesenchymal stem cells, after induced differentiation by 5-azacytidine, expression of cardiomyocyte-specific proteins in the cardiomyocyte-like cells is detected, and besides, the expression is significantly increased, indicating that the Rspo1 promotes the differentiation of the bone marrow mesenchymal stem cells into the cardiomyocyte-like cells and a conversion rate of the cardiomyocyte-like cells is as high as 53%.

The invention discloses a rat macrophage culture method. The method comprises the following steps: killing young SD rats which are born for 3 weeks by a neck removal method, soaking the rats in 75% alcohol for disinfection for 3 minutes, transferring the rats into an ultra-clean workbench for operation after disinfection is completed, cutting posterior limb skin, removing leg muscles, exposing thighbones, stripping external muscles of the thighbones cleanly, and cutting off the thighbones; cutting off femoral heads at the two ends of the thighbones, sucking a certain amount of MEM-alpha basalculture medium liquid by using an injector, blowing out bone marrow, and flushing the bone marrow cleanly in the blowing process; blowing and beating the bone marrow for 30 times by using a pasteur pipette to prepare a single-cell suspension, performing centrifuging by using a centrifuge, collecting cells, and discarding the supernatant; and adding erythrocyte lysate. According to the invention, the rat bone marrow is subjected to primary centrifugal separation, erythrocyte lysis, secondary centrifugal separation and culture by using a special culture medium for macrophages, so that the cultured macrophages are relatively high in purity, the culture speed is relative high, and the research requirements are met.

The invention relates to a method for inducing minimal embryonic stem cells to differentiate into cardiomyocytes. The method includes the following steps: (1) obtaining CD45-Lin-CD106+cells (minimal embryonic stem cells) in rat bone marrow through flow cytometry sorting; (2) inducing, by 5-azacytidine, the minimal embryonic stem cells to differentiate; and (3) through qRT-PCR and immunofluorescence, detecting myocardial protein related genes, and validating the process that the 5-azacytidine induces the minimal embryonic stem cells to differentiate. The purpose of the method is to isolate andculture the minimal embryonic stem cells from rat bone marrow tissue; through the adoption of the 5-azacytidine to induce and differentiate, biological characteristics and directed differentiation characteristics can be observed, so that a convenient and fast approach for cell transplantation treatment of clinical bradyarrhythmia can be found. Validation results are reasonable and accurate, and experimental processes are precise and cautious.

The invention discloses a rat bone marrow tumour cell NSO and a preparation method and an application thereof. The rat bone marrow tumour cell NSO can grow in a serum-free and fat-free culture substrate, the preservation number of one strain (NSO-Ch1) is CGMCC No.2127. If the rat bone marrow tumour cell NSO of the invention, which can grow in a serum-free and fat-free culture substrate, is used to express foreign protein, cell culture efficiency and expression efficiency can be ensured, and other animal source ingredients and unsafe factors can be avoided; thus, the invention can effectively lower purification cost and loss and improve the yield and profitability of biological products.

The invention discloses a method for alkaline phosphatase to induce gelation and biomimetic mineralization of silk protein solution. The invention introduces a small molecule polypeptide that is sensitive to ALP and has good biocompatibility and self-assembly properties as a gelling factor The precursor, which can remove the phosphate group on the molecule under the catalysis of ALP to generate NY, triggers supramolecular self-assembly, thereby synergistically inducing SF co-self-assembly, and finally leads to the rapid formation of SF hydrogel. The ALP encapsulated in the SF‑NY hydrogel network still maintains its catalytic activity and induces the formation of apatite minerals in the gel by catalyzing β‑glycerophosphate to release free phosphate ions. Due to the mild gelation process and the formation of apatite minerals in the gel matrix, the biomimetic mineralized SF gel could not only be used as a biomimetic scaffold to promote the adhesion of rat BMSCs in vitro, Proliferation and osteogenic differentiation, and can also promote natural healing of femoral defects in a rat model.

The invention discloses a magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2 and a preparation method thereof. The method for preparing the magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2 comprises the steps of: firstly preparing hydrophobic monodisperse Fe3O4 nano-particles by adopting a chemical oil-phase high-temperature method, and modifying the surfaces of the hydrophobic Fe3O4 nano-particles to ensure that the hydrophobic Fe3O4 nano-particles are dispersed in a water phase; preparing luminescent CdTe quantum dots of which the surfaces are provided with carboxyl groups, and precipitating the luminescent CdTe quantum dots on the surfaces of the magnetic Fe3O4 nano-particles through the co-precipitation; then utilizing ligand exchange to modify a silane coupling agent on the surfaces of the luminescent CdTe quantum dots; and finally forming an outermost SiO2 coating layer through silane or silicon ester hydrolysis. The diameter of the magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2 is between 30 and 50nm; the magnetic luminescent composite nano-particle Fe3O4 / CdTe / SiO2has double functions of magnetism and fluorescence at the same time, has strong and durable fluorescence intensity after labeling rat bone marrow-derived mesenchymal stem cells, and apparently reduces cellular magnetic resonance signals. The particle has broad application prospect in the fields such as biological labeling, bioseparation and the like.

The invention relates to the technical field of biological implant materials, in particular to a titanium implant with a nanometer bionic three-dimensional porous titanium trabecula structure and a preparation method and application thereof.The method comprises the steps that in the presence of electrolyte, a titanium substrate M0 is subjected to anodic oxidation, and a titanium substrate M1 with a deposited titanium dioxide nanotube array layer is obtained; removing the titanium dioxide nanotube loaded on the titanium substrate M1 to obtain a titanium substrate M2 loaded with a nano bowl structure; the titanium matrix M2 is placed in an alkaline solution to be subjected to etching treatment, and the titanium implant with the nanometer bionic three-dimensional porous titanium trabecular structure is obtained. The titanium implant with the nano bionic three-dimensional porous titanium trabecula structure prepared on the basis of the method provided by the invention can promote adhesion, proliferation and differentiation capacities of rat bone marrow stromal stem cells in vitro and promote osseointegration of the titanium implant in vivo.

The invention provides a immortal stromal cell systme of rat's marrow and preparation thereof. Immortal stromal cell of rat's marrow can be obtained in short time by the inventive method. The inventive hTERT immortal stromal cell systme of rat's marrow is stable, can be sequentially passed through generations and keep diploid karyotype and characteristics of normal stromal cell of rat's marrow, has extremely strong proliferation ability, and realize immortalization. Immortal stromal cell system of rat's marrow established by the inventive method can conveniently and quickly obtain large amount of stromal cell of rat's marrow with diploid karyotype. This no doubt has great promotion to research of introduction and bone marrow stromal cell in vitro and in vivo.

The invention provides a polylactic acid composite biological material as well as a preparation method and application thereof. The polylactic acid composite biological material comprises a compound of which the surface is coated with a silane coupling agent magnesium silicate and polylactic acid. The polylactic acid composite biological material can obviously promote adhesion and proliferation of rat bone marrow mesenchymal stem cells and activity expression of alkaline phosphatase, and is beneficial to promoting in-vivo osteogenesis. The preparation method is simple and convenient to operate; wherein the prepared polylactic acid composite biological material and the 3D biological printing scaffold have excellent in-vitro degradability and biological activity, can release silicon and magnesium ions, are beneficial to cell osteogenic differentiation and can be applied to tissue engineering.

The invention relates to a method for improving the efficiency of differentiating rat bone marrow mesenchymal stem cells into myocardial cells, which comprises the following steps: (1) constructing alentiviral interference vector G9a-shRNA, and transfecting the G9a-shRNA to the rat bone marrow mesenchymal stem cells; and (2) inducing the bone marrow mesenchymal stem cells transfected with the interference vector through 5-azacytidine, and detecting the positive rate of troponin positive cells by flow cytometry on the 21st day. According to the invention, in the process of inducing the bone marrow mesenchymal stem cells to differentiate into the myocardial cells by the 5-azacytidine, the H3K9me2 methylase G9a is knocked down, and the final induction efficiency is observed, so that a convenient and rapid way is expected to be found in the process of differentiating the bone marrow mesenchymal stem cells into the myocardial cells. According to the method, the rat bone marrow mesenchymalstem cells are transfected by adopting FuGENE (Promega) lipidosome mediated G9a-shRNA lentivirus recombinant plasmids to reduce the expression of H3K9me2, so that the induction efficiency is improved,the verification result is reasonable and accurate, and the experimental process is rigorous and rigorous.

The invention discloses a rat bone marrow derived mast cell culture method. The culture method comprises the following steps: killing SD young rats born for 3-5 weeks by a neck-removing method, soaking the SD young rats in 75% alcohol for disinfection for 3 minutes, then transferring the rats into an ultra-clean workbench for operation, cutting off hind limb skin, removing leg muscles, exposing femur, peeling off external muscles of the femur, and cutting off the femur; cutting off femoral heads at the two ends of the femur, sucking a DMEM / F12 basal culture medium by using an injector, blowingand beating out femoral marrow, and flushing the marrow cleanly in the blowing and beating process; blowing and beating the bone marrow for 30 times by using a Pasteur pipette to prepare a single-cell suspension, performing centrifuging by using a centrifuge, collecting cells, and discarding supernatant; and adding red blood cell lysate and carrying out lysis at 4 DEG C for 3 min. According to the method, the mast cell induction culture medium and the mast cell maturation promoting culture medium are used for culture and multiple times of centrifugal separation, so that the cultured mast cells are relatively high in purity, and the culture period is greatly shortened.

The invention discloses a method for constructing a rat bone mesenchymal stem cells (BMSCs) cell strain for expressing c-met protein through regulation of doxycycline. The method comprises the steps that an rtTA gene lentiviral expression vector is constructed and subjected to transfection into BMSCs so that a BMSCs cell strain (a Tet-On Advanced BMSCs cell strain) for stably and efficiently expressing rtTA protein can be obtained; then, a Len-TRE min CMV-c-met / ALB-GFP lentiviral vector is constructed and subjected to transfection into the Tet-On Advanced BMSCs, and a flow cytometry is used for screening GFP positive BMSCs; and finally the GFP (+) Tet-on-c-met BMSCs cell strain is obtained. The doxycycline can be added into the obtained BMSCs cell strain to induce the obtained BMSCs to express the c-met protein, and the method has important significance for study on blood doping of the cell strain for treating hepatic failures, the directional homing and differentiation capacities of the BMSCs along with the HGF concentration and the dose-effect relationship between the capacities and the HGF / c-met axis expression level.

The invention discloses an SD rat-derived bone marrow mesenchymal stem cell induction medium and an induction method. The induction medium is composed of α-MEM medium and additives. In terms of final concentration, the additives are 5-100ng / ml Composition of transforming growth factor-alpha, 10-200ng / ml regulatory protein beta 1, 100-3000nM trehalose and 50-5000nM dibutyryl cyclic adenosine monophosphate. The SD rat-derived bone marrow mesenchymal stem cell induction medium and induction method of the present invention can significantly increase the secretion of BDNF, HGF, VEGF, GDNF and other factors of rat bone marrow mesenchymal stem cells, and the types of secreted factors are also relatively abundant and It can be secreted stably for a long time; the method of cytokine induction avoids the risks and uncertainties brought about by other methods such as genetic modification, and is more suitable for industrialization and clinical application.

The invention provides a ZIF-8-loaded Janus electrospun fibrous membrane as well as a preparation method and application thereof, and belongs to the technical field of dental restoration materials. The method comprises the following steps: mixing ZIF-8 with a polyvinylpyrrolidone solution, mixing the mixed solution with a poly (epsilon-caprolactone) solution in an isovolumetric manner, and obtaining the Janus electrospun fiber membrane by utilizing a phase separation phenomenon between the polyvinylpyrrolidone solution and the poly (epsilon-caprolactone) solution through a single-shaft electrospinning technology. The invention further provides the Janus electrospun fiber membrane of the ZIF-8, which is obtained by the preparation method. According to the Janus electrospun fibrous membrane, slow release of ZIF-8 is achieved, proliferation of bacteria can be inhibited, osteogenic differentiation of rat bone marrow mesenchymal stem cells is effectively promoted, and the Janus electrospun fibrous membrane has a good treatment effect on periodontal defects caused by periodontitis.

The invention belongs to the technical field of medicine and relates to uses of various traditional Chinese medicine mixed compounds, in particular to application of an effective part group of yang-tonifying recuperating decoction in inducing mesenchymal stem cells to differentiate into nerve cells.The effective part group includes effective parts extracted from medicinal materials, including glucoside compounds, total alkaloids, Angelica sinensis polysaccharide, carthamin yellow and naringenin.Researches show that this effective part group can induce in vitro rat mesenchymal stem cells to differentiate into neuron-like cells and is applicable to drugs for treating nervous system diseases.