49 results about "Orthotopic transplantation" patented technology

The invention relates to a lentivirus vector expression system of luciferase and an application thereof. Particularly speaking, in the invention a luciferase lentivirus expression system with FG12-Luc2 as core plasmid is successfully constructed, and a PVTT-1 cell line from tumor thrombus tissue of liver cancer patients is established by the system. Through the lentivirus expression system, liver cancer tissue is successfully marked; and by experimental methods such as subcutaneous transplantation and orthotopic liver cancer transplantation, a mouse orthotopic transplantation liver cancer metastasis model is established which is suitable for serial passage, is applicable to in-vivo imaging, and can realize real-time monitoring of the growth and metastasis conditions of tumors in the body.

The invention belongs to the biological medicine field, and discloses a targeted boron preparation and a preparation method and application. The target boron preparation comprises polyamidoamine-amine dendrimers, epidermal growth factor receptor antibody and polyhedral boranes, the polyamide amine dendrimers are connected with the epidermal growth factor receptor antibody, and are internally loaded with the polyhedral boranes. The targeted boron preparation is high in boron content, can meet the requirements of BNCT (boron neutron capture therapy) on the boron atom dose, is well targeted to tumor cells, has good stability and low cell toxicity, and can be efficiently uptaken by human glioma U87MG cells of high surface expression EGFR (epidermal growth factor receptor). The target boron preparation can effectively improve the content of boron in orthotopic transplantation tumor nude mice tumor tissues, can significantly prolong orthotopic transplantation tumor nude mice lifetime by neutron irradiation, and is suitable for boron neutron capture therapy for glioma. The preparation method of the targeted boron preparation has the advantages of simple operation, and the prepared targeted boron preparation has good stability.

The invention pertains to the field of biomedical technology, in particular to an orthotopic transplanted mouse liver cancer model which takes the research of the occurrence and development mechanisms of liver cancer and the development of anti-tumor drugs as the purposes, and the preparation method and the application thereof. The H22 cell strain of stable transfection EGFP is injected into the BALB / c mouse peritoneal cavity for amplification; the ascites is extracted from the H22 ascites cancer mouse peritoneal cavity, then cell separation is carried out and cell suspension is prepared; the mouse peritoneal cavity is opened, and the cell suspension is extracted by a micro-syringe to be injected into the mouse liver; a cotton bud with 75 percent alcohol is pressed on the needle hole till the surface of the liver does not bleed any more, and the needle hole is sealed by bonding agent; then the liver surface is washed by physiological saline, and layer separation is carried out and the peritoneal cavity is closed. The results show that the occurrence rate of mouse liver cancer is 100 percent and the natural remission rate is zero. The model is an ideal orthotopic liver cancer model which can be used in the research of the development and metastasis mechanisms of orthotopic liver cancer and can also be applicable to the development of anti-tumor drugs, the experimental treatment and diagnosis research of liver cancer.

The invention discloses a manufacturing method of a biological tooth root bracket material. The method comprises the following steps of: flushing teeth with phosphate buffered saline (PBS) in a sterile state for three times; removing dental crown parts by grinding with a carborundum disc, and keeping tooth root parts; pulling pulp tissues out of the tooth roots and repeatedly flushing the inside of root canals with physiological saline; gradually expanding the inside of teeth with a dental expander and repeatedly flushing the inside of the root canals with 5 to 17 percent of ethylene diamine tetraacetic acid (EDTA) solution; grinding outer side tissues of the tooth roots by grinding with a dental turbine and constructing a biological tooth root profile as required so as to obtain a bracket with a specific shape; preserving the teeth in the PBS and treating in an ultrasonic oscillator for 3 to 8 times; performing stepped demineralization on a dentine with EDTA with different concentrations in a magnetic stirrer with the rotating speed of between 100 and 500 revolutions per minute so as to obtain a dentine substrate; and preserving the dentine substrate in double-antibody-containing PBS. The biological tooth root bracket material has a tooth root profile, the thickness of 1mm to 3mm, the length of 1cm to 2.5cm and certain tooth inducing capability, comprises a large number of proteins and factors related to the manufacturing of teeth, can release the proteins and factors continuously, can better solve the problem of induction micro-environment, and contributes to application to tooth socket intra-alveolar orthotopic transplantation.

The invention provides a method for establishing orthotopic transplantation model of colorectal cancer, which comprises a) selecting and culturing colon adenocarcinoma cell line and preparing cell suspension; b) performing colostomy on living experimental animal; c) and inoculating the above colon adenocarcinoma cell suspension to submucosal stroma to allow natural growth. The biological traits such as route of metastasis of the transplanted adenocarcinoma cells are similar to that of the primary colon adenocarcinoma cell. The invention is advantageous in that the model is easy to be established, the transplanted colon adenocarcinoma locates at body surface so that it can be sampled and observed conveniently to meet the requirement for dynamic research, and the consumption of experimental animals can be reduced.

The invention discloses a method for establishing a green fluorescence BALB / c nude mouse model. The method includes: selecting female C57BL / 6-Tg (CAG-EGFP) mice and male Foxn1nu mice to perform hybrid to acquire F1 child generation hairy fluorescence mice, selecting the female mice, selecting the female mice having sexual maturation and the parental generation male Foxn1nu mice to perform mating (backcross) to acquire an F2 child generation; generating segregation of characters of the F2 child generation, selecting female hairy fluorescence mice and the parental generation male Foxn1nu mice to perform backcross, and completing a backcross cycle; performing the backcross cycle for ten times to acquire an F10 child generation, and establishing the green fluorescence BALB / c nude mouse model. The invention also discloses application of an animal model established by the method to a brain tumor transplanting model. The method uses a homogenous introduction method to establish the new strain model mice which can stably express EGFP, and can lay a firm foundation for human glioma orthotopic transplantation tracking and research on glioma and a micro environment thereof.

The invention provides a method for establishing an inflammation-based pancreatic cancer animal model, which comprises the following steps: 1. constructing a pLVX-luciferase-IRES-ZsGreen1 plasmid; 2. establishing a slow-virus-mediated luciferase overexpression system; 3. establishing a pancreatic cancer cell line for expressing luciferase; 4. establishing a nude mouse pancreatic gland orthotopic transplantation tumor model; 5. establishing an inflammation model; and 6. establishing the inflammation-based pancreatic cancer animal model in the nude mouse body by continuing breeding the nude mouse for 30 days. The pancreatic cancer animal model established by the method can catalyze the substrate to emit light by using the luciferase, and the size of the tumor can be evaluated after the light is detected by the corresponding instrument and equipment.

The invention discloses a transplantation tumor model of colorectal tumor for medical scientific research, which is produced by improving the orthotopic transplantation tumor model of colorectal tumor in nude mice. The invention aims to overcome the defect that the orthotopic transplantation tumor model of colorectal tumor has high intestinal obstruction incidence rate and short model animal survival time and is not suitable for long-time experiments. In the invention, the prepared tumor is transplanted close to the blood vessel in mesocolon. The method comprises the following steps: (1) preparation of tumor: injecting single cell suspension of the tumor cell to subcutis of nude mice and preparing the source of the transplantation tumor; (2) transplanting the prepared tumor close to the blood vessel in mesenterium of nude mice but not the serosa of the intestinal tube; and (3) raising the nude mice in an aseptic laminar flow room, controlling the room temperature to be (25+ / -1) DEG C and relative humidity to be 40-50% and continuously observing the nude mice. The animal model built by the invention can reproduce the natural clinical pathology process of human colorectal tumor, can not suffer from obstruction of the lumen in the digestive tract in the short term and has transfer mode similar to that of the patients.

The invention provides a lymph gland targeted metastatic human hepatoma cell strain. The mother cell is a red fluorescent protein-expressed hepatoma cell HCCLM3-R. The human hepatoma cell strain is characterized in that the cell is named as lymph gland targeted metastatic human hepatoma cell strain HCCLM3-R-LnM1 according to the classified nomenclature with a preservation number of 3645, and has the following biological characteristics: the cell is polygonal epithelioid and is in adherent growth and loss of contact growth inhibition, is of a hypo-triploid karyotype, and has 50-58 of chromosomes each of which secretes AFP proteins. The establishing method comprises the following steps: using subcutaneous and liver orthotopic transplantation tumor models of a nude mouse to carry out spontaneous lymphatic metastasis screening, and then carrying out in-vitro cloning, amplification and purification on the lymphatic metastasis human hepatoma cell. The invention has wide application prospects and potential social and economic values in the basic research on hepatoma and the pre-clinic research on medicament.

Human breast carcinoma SK-BR-3 cell strain as source cell for building train adapted by the invention is continuously inoculated on female NOD / SCID mouse mammary gland fat pad and filtered in pressure of low dose chemotherapy drug tail intravenous injection to build human breast carcinoma cell line SK-3rd rich in tumour breast carcinoma stem cell. The proper of knub stem cell in the cell is increased greatly comparing with non NOD / SCID vivo passage source cell. The formation rate of saccule and the number of cell in the saccule are increased greatly in case of cell suspension culture. The ratio of undifferentiation cancer cell is reduced but is higher evidently than source cell in process of SK-3rd saccule cell inducing differentiation. The saccule cell formed from SK-3rd cell has carcinoma stem cell phenotype which gradually dies out in process of differentiation. NOD / SCID mouse orthotopic transplantation has high tumour-forming rate, high long-distance transferring rate, short tumour-forming periodic time and less number of cells needed by tumour-forming. The invention is an ideal mould for studying breast carcinoma stem cell and has wide applications foreground for disclosing generation mechanism of breast carcinoma and neoplastic treat.

The invention relates to a method for establishing a model of fluorescence orthotopic transplantation of subcutaneous metastasis organ cancers, which is characterized in that human colon adenocarcinoma cell strains or cancer cell strain green fluorescent proteins (GFP) of corresponding organs are selected; GFP plasmids carry out transfection on cancer cell strains of the organs via retrovirus pLPCX so that the cancer cell strains of the organs contain GFP expression plasmids; a GFP gene is connected with a pLPCX vector via a T4 ligase; a GFP-pLPCX vector is transfected to a PT67 package cell to obtain a Lovo cell; nude mice are selected and subjected to subcutaneous inoculation with Lovo cell suspension; tumors can be formed in about 4 weeks; orthotopic transplantation is carried out after green fluorescence is observed; a cut is opened on the skin and peritoneum; about 1cm cecum of the nude mice is taken out of the peritoneum; the gap between the cecum and the peritoneum is sutured and the cecum is transferred subcutaneously; the cut cecum is subjected to serosa; two fluorescence-carrying tumors with diameter of about 1mm are transplanted to the cecum via the suture and the skin is closed.

The invention provides a method for constructing engineering arterial blood vessel in vivo by taking melt-spinning fiber as a skeleton. The method comprises the following steps: firstly preparing a melt spinning and medical silica gel tube composite mandrel, and then preparing the in-vivo engineering arterial blood vessel taking melt-spinning fiber as a skeleton by taking subcutaneous part of an animal as a bioreactor; the specific steps are as follows: 1) preparing the melt spinning and medical silica gel tube composite mandrel; 2) preparing the in-vivo engineering arterial blood vessel taking melt-spinning fiber as a skeleton by taking subcutaneous part of an animal as a bioreactor; and 3) performing in-situ transplanting of the in-vivo engineering arterial blood vessel so as to replace lesion blood vessels. The method has the advantages that the in-vivo engineering arterial blood vessel has good biocompatibility, all the components are originated from autologous tissues, and therefore, the artificial blood vessel is non-toxic, free from immunogenicity, and incapable of causing inflammation; due to the existence of melt spinning fiber, the prepared in-vivo engineering blood vessel has good mechanical strength, tenacity and compliance, and is suitable for surgical sewing operation.

The invention discloses a modeling device for establishing a bladder cancer orthotopic transplantation tumor animal model, solving the defects of long surgical time, complex operation, large animal bladder mucosa hurt and the like of the traditional modeling method. The modeling device comprises a needle seat fixedly arranged at the front end and a sleeve base provided with a cavity, wherein the front end of the sleeve base is fixedly provided with a soft sleeve communicated with the cavity, the front end of the needle base is sleeved and inserted in the cavity of the sleeve base and positioned by using a positioning baffle ring arranged on the needle base, a needle core on the needle base is sleeved and inserted in the soft sleeve of the sleeve base, and the length of the needle point ofthe needle core, exposed out of the end part of the soft sleeve, is 0.5-1.0mm, a break angle of 5-7 DEG is arranged at a place of the needle core, which is at a 1.5-2.0cm distance of the needle point, and a marking groove is arranged on the positioning baffle ring. The modeling device has the advantages of skilled design, simple structure and easiness in operation, greatly shortens the surgical time, and has little damage to the animal bladder mucosa and a tumor formation rate of reaching 100 percent.

The invention relates to the technical field of biology, in particular to a building method of a human choriocarcinoma naked mouse orthotopic transplantation tumor model. The building method includessteps: S1, culturing human choriocarcinoma JAR cells; S2, selecting a female naked mouse, and injecting 200ul of a human choriocarcinoma JAR cell suspension to a right-side inguinal position; removingtumor tissue when tumor grows to 2cm3 in size; S3, in a sterile condition, using sterile normal saline to rinse choriocarcinoma tissue, cutting into tissue blocks of 1mm3, and putting into the sterile normal saline for standby use; selecting a female naked mouse, performing general anesthesia, transplanting the choriocarcinoma tissue blocks into corporis uteri cavum, suturing layer by layer, andclosing abdomen; continuing to feed after operation, and allowing free intake, in this way, the human choriocarcinoma naked mouse orthotopic transplantation tumor model is built in the body of the naked mouse. By simulating tumor cell growth microenvironment and conforming to clinical genesis and development of tumor, a good foundation is laid for further studying pathogenesis and clinical treatment of human choriocarcinoma.

The invention relates to a high metastasis human breast carcinoma cell subline MDA-MB-231sci, which has biological and pathological features and special metastasis phenotypes. The biological features are as follows: the MDA-MB-231sci cells are typical polygonal epithelioid cells, have rich cytoplasm and big and round karyon, grow adherent to the wall with almost uniform size and are closely arranged with clear boundary. The pathological features are as follows: a tumor cell of a nude mouse in vivo metastasis part has the similar structure with the subcutaneous tumor cell, short metastasis time and high metastasis rate, wherein the metastasis time is shortened to 26-42 days; a lung metastasis rate is 100%, meanwhile, a part of axillary lymph node metastasis of an animal is performed. An establishing method for the high metastasis human breast carcinoma cell line comprises the following steps of: (1) establishing a human breast carcinoma orthotopic transplantation tumor model; (2) sieving an orthotopic transplantation lung targeted metastasis model; and (3) establishing and passaging a human breast and lung carcinomas targeted high metastasis cell line. In the invention, the success rate is improved by sieving surgical excision tumor of each generation in vivo and corresponding breast.

The invention discloses a novel mouse in-situ pleural mesothelioma model and an establishment method thereof. The establishment method comprises the following steps of 1, taking 90 BLAB / C nude mice as an experimental group and 30 control groups, and uniformly putting the experimental group and the control groups into an incubator for normal culture; 2, under the condition that BLAB / C nude mice breathe, heartbeat and the like are normal, anesthetizing the mice and fixing the mice on an animal operating table in a supine position for experiment; and 3, at the intersection point of the left rib 5 / 6 intercostal space and the left chest wall anterior axillary line of the mouse, using microscopic forceps to lift and pull the skin at the intersection point, then using a 22G sterile sharp needle head to break the skin at the intersection point, then using a 30G sterile blunt pillow to break the wall layer pleura along the broken opening. The tumor transplantation method provided by the invention is applicable to orthotopic transplantation of pleural tumors. The growth mode, the invasion ability and the tumor metastasis effect of the pleural tumor can be restored to the maximum extent, and a good mouse model is constructed for studying the pleural tumor.

The invention provides a construction method of a malignant neurinoma xenograft mouse model. The method comprises the following steps of: resuscitating a tumor tissue block of a malignant neurinoma patient stored in a liquid nitrogen environment; and carrying out xenogeneic subcutaneous transplantation on the resuscitated tumor tissue block of the patient into immunodeficient mouse bodies under asterile condition. The xenogeneic subcutaneous transplantation comprises the following steps: selecting immunodeficient mice with the age of 2-6 weeks, wherein the sex of the mice is kept consistent with that of the patient; soaking the resuscitated tumor tissue block of the patient in sterile incubation liquid containing fetal calf serum; fixing the mice after anesthesia; implanting the tumor tissue block of the patient under the skin of the mice; suturing incision; and raising the mice to enable tumors in the body of the mice to grow. The method also includes: performing xenogeneic orthotopic transplantation on the cryopreserved and resuscitated tumor tissue block of the patient into immunodeficient mice. According to the invention, the xenograft mice are used for replacing clinical patients to screen different combined medicines, so that the most appropriate medicine is selected, the effectiveness is greatly improved, and the side effects of the medicines are reduced.

The invention discloses a construction method of a glioma cell line suitable for two-photon living imaging. The construction method comprises the following steps: constructing a glioma cell in-situ transplantation animal model through glioma cells, and researching the dynamic development change of the glioma cells in the brain of a mouse by virtue of a two-photon living body imaging technology. The GL261-ZsGreen glioma cells obtained through transformation can be successfully used for constructing a glioma in-situ transplantation living animal model, the positions of tumor cells can be clearlyobserved in the imaging observation process, cell-level examination can be accurately performed when the GL261-ZsGreen glioma cells are applied to the research field, the glioma boundary can be obviously distinguished, and a potential means is provided for effectively and accurately excising tumors. Each proliferated GL261-ZsGreen cancer cell is provided with a green fluorescence label, so that single cancer cells subjected to dispersive metastasis can be accurately positioned, and a possible scheme is provided for further researching the diffusion of cancer cells and the accurate treatment of glioma.

The invention relates to a device for establishing a mouse hepatic cancer orthotopic transplantation tumor model, this device being accurate to position and practical and convenient to use during injection to a liver of a mouse. The device comprises a positioning plate A and a positioning plate B, the positioning plate A includes a front plane A, a rear plane and a right plate, a groove A is connected between the front plane A and the rear plane, and the front face of the groove A is provided with an injecting hole; the positioning plate B comprises a front plane B, a rear bevel and a left plate, a groove B is connected between the front plane B and the rear bevel, and the front face of the groove B is provided with an injecting hole; the positioning plate A and the positioning plate B are connected through groove inserting to form an injection positioning plate, and after inserting, the positioning plate B is positioned above the positioning plate A. The injection positioning plate is accurate to position and easy to operate, reproducibility of experiments is guaranteed, model success rate is further increased, and the device is applicable to experimental operations in various prevention and treatment studies on various liver cancer, such as establishment of animal models and mouse liver orthotopic injection.

The invention discloses a preparation method of mouse ECMs (Extracellular Matrixes), obtained mouse ECMs and a mouse ovarium in-vivo regeneration method. The method comprises the following steps of 1, performing treatment for different time by using detergents with different concentrations, removing cells of mouse tissue organs, and preparing the ECMs; 2, performing histological analysis and biochemical analysis on the prepared ECMs so as to determine the optimum treatment condition; and 3, performing orthotopic transplantation on the ECMs subjected to cell removal treatment, recording the development condition of the ECMs and the ovarium regeneration efficiency, and further, verifying the regenerated ovarium tissues by a histology method, an immunohistochemistry method, a PCR (Polymerase Chain Reaction) method and the like. In addition, the effect that the tissue scaffolds (ECM) from different sources can also promote the ovarium regeneration is also proved. By using the method provided by the invention, the ovarium is successfully regenerated for the first time; and the further solving method is provided for the problem of female apogeny due to ovarium dysfunction. The method provided by the invention has the advantages of simplicity, convenience, low cost, low experiment condition requirements and high operability.

Described herein are methods and compositions for regulation of the p53 pathway, providing intrinsic “sternness” allowing for their efficient in vitro expansion following isolation. Pharmacologic approaches to modulate p53 signaling supports expansion of stem cells, including greater clonal expansion of lung stem cells when compared to use of other small molecules such as ROCK inhibitor Y27632 alone. Effects of combined treatment with Pifithrin-α and Y27632 are additive. The current invention involves use of drugs that target the p53 pathway to reversibly regulate stem cell expansion in vitro for banking of stem cells and for pre-conditioning of stem cells prior to orthotopic transplantation.

The invention discloses a traditional Chinese medicine composition for adjuvant treatment of a liver cancer. The traditional Chinese medicine composition is prepared from 10-20 parts of astragali radix, 5-15 parts of curcuma zedoary, 10-30 parts of manyleaf paris rhizome, 10-20 parts of hairyvein agrimony, 5-10 parts of radix asparagi, 5-10 parts of coix seeds, 15-30 parts of ardisiae japonica, 15-30 parts of oldenlandia diffusa, 15-30 parts of rhizoma pinelliae and 15-30 parts of chinaroot greenbrier, 10-40 parts of ganoderma lucidum and 15-30 parts of Chinese actinidia roots. It is shown according to experimental results that the traditional Chinese medicine composition combined with the chemotherapeutic drug cisplatin can obviously inhibit the growth and metastasis of liver cancer orthotopic transplanted tumors, can reduce the toxic and side effects of the chemotherapeutic drug, prevent the damage of the cisplatin to immune organs and improve the inhibition effect of the cisplatin on bone marrow. The traditional Chinese medicine composition combined with the chemotherapeutic drug cisplatin induces apoptosis of the human hepatoma cells HepG2, which indicates that the combined useof the traditional Chinese medicine composition and the chemotherapeutic drug cisplatin can produce effect-enhancing and toxicity-reducing action, and the traditional Chinese medicine can be used asan adjuvant radio therapy and chemotherapy medicine for clinically treating tumors.

The invention discloses the application of an lnc-ZNF100-5 functional expression inhibitor in the preparation of a drug for treating breast cancer, and belongs to the technical field of biomedicine. The lnc-ZNF100-5 functional expression inhibitor is RNAi, shRNA lentivirus, sgRNA, ZFN, TALEN element or homologous recombination vector. In the present invention, the expression of lnc-ZNF100-5 in breast cancer cells is significantly increased, which can be used for the detection of breast cancer, and an in vitro lnc-ZNF100-5 knockout breast cancer cell model and an in vivo lnc-ZNF100-5 knockout breast cancer cell model are established. A series of experimental studies on cancer orthotopic xenograft models have demonstrated that lnc‑ZNF100‑5 can significantly inhibit the proliferation of breast cancer cells and significantly promote the apoptosis of breast cancer cells. Knocking out lnc‑ZNF100‑5 can be used for prevention or treatment Breast cancer, so that lnc-ZNF100-5 functional expression inhibitor can be prepared to treat breast cancer.

The invention discloses a method for building a triple negative breast cancer living tumor tissue mouse orthotopic transplantation model. The method comprises the following steps of (1) subcutaneouslyembedding an estradiol sustained-release tablet in the neck of an immunodeficient mouse; (2) selecting triple negative breast cancer living tumor tissues of a clinical triple negative breast cancer patient through an operation, and then cutting the triple negative breast cancer living tumor tissues into inoculation tumor blocks with the volume being 4 to 15 mm<3>; (3) performing orthotopic transplantation on the inoculation tumor blocks into a mammary gland fat pad of the immunodeficient mouse; and (4) raising the immunodeficient mouse for 30 to 80 days, and building the triple negative breast cancer living tumor tissue mouse orthotopic transplantation model. The method has the advantages that the estrogen level in a female mouse model body can be improved; and the growth requirement of the triple negative breast cancer tissues is met, so that the tumor nodule emergence time is early, the model building time is shortened, the tumor forming rate is improved by nearly 4 times, and the model building method is simple and feasible.

The invention relates to the technical field of medicines, and particularly discloses application of berberine hydrochloride in restraining the pancreatic cancer. After the interference of the berberine hydrochloride, the growth speed of the pancreatic cancer orthotopic transplantation tumor is obviously reduced, and the tumor size is obviously reduced; meanwhile, the berberine hydrochloride has the remarkable restraining effect on the pancreatic cancer cell proliferation. Specially, we find that the berberine hydrochloride has the good restraining effect on the pancreatic cancer under the lowdosage. In addition, the berberine hydrochloride is rich in source, low in price and high in safety, and therefore the cost for treating the pancreatic cancer can be reduced and the toxic and side effects caused by treatment can be reduced by using the berberine hydrochloride as the pancreatic cancer treatment means.

The invention discloses the use of combination of cytokine and hypoglycemic in treatment of diabetes, wherein on the basis of using conventional hypoglycemic medicament for diabetes treatment, bone marrow hemapoietic stem cell mobilizing agent-recombinant human granular leukocyte colony stimulating factor (GCSF) is integrated, thus the damaged pancreatic island tissues can be renovated through mobilizing the orthotopic transplantation on pancreatic island positions inside stem cells, and achieving the goal of diabetes treatment.