33results about How to "Non-tumorigenic" patented technology

The invention relates to umbilical cord mesenchymal stem cell injection and a preparation method and application thereof. The injection comprises umbilical cord mesenchymal stem cells and mixed solution; the mixed solution comprises balanced electrolyte solution, hydroxyethyl starch, adenosine disodium triphosphate-magnesium chloride, dimethyl sulfoxide and human serum albumin. The preparation method comprises the following steps: (1) fully and evenly mixing the balanced electrolyte solution, the hydroxyethyl starch and the human serum albumin, and performing filtration sterilization; (2) adding the umbilical cord mesenchymal stem cells; (3) adding the dimethyl sulfoxide; (4) performing split charging and frozen preservation to finish preparation; the invention also provides the application of the injection in the aspect of drugs for treating chronic ischemic heart diseases. The injection disclosed by the invention has a very good resuscitation effect after the frozen preservation, can be directly injected after being unfrozen and resuscitated, and has a remarkable effect on treatment on the chronic ischemic heart diseases.

Provided herein are mesenchymal stem cells which have been modified by the introduction of polynucleotides encoding for a mammalian Cdk1 protein. These cells do not sense in culture and are non-tumorigenic thereby providing an ongoing source of cells as well as conditioned medium. The conditioned medium from these cells can be used for tissue repair. Also provided is a method of modifying mesenchymal stem cells which can be continuously propagated in culture and are non-tumorigenic.

The invention discloses a method of induced neural stem cells and a composition. Through the method and the composition, peripheral blood mononuclear cells can be induced to form neural stem cells. The neural stem cells can express neural stem cell related genes, and can disintegrate out nerve cells, astroglia cells and oligodendrocyte. Dopaminergic nerve precursor cells formed through in vitro induction and disintegration of the neural stem cells are transplanted into the striatum corpora of a PD mouse model, no tumors are formed, the ethology of a mouse Parkinson 's disease model can be improved, and the process of the Parkinson 's disease can be delayed. The neural stem cell inducing and disintegrating method provided by the invention is simple and quick to operate, small in traumatic occlusion and good in safety, and is hopefully used for treating the Parkinson 's disease.

The invention relates to the technical field of stem cells and especially relates to a placenta mesenchymal stem cell injection, a preparation method and application thereof. The placenta mesenchymal stem cell injection comprises placenta mesenchymal stem cells, vitamin C, dimethyl sulfoxide (DMSO), fetal calf serum (FBS), human albumin, rhizoma panacis majoris saponin and solvent. The placenta mesenchymal stem cell injection prepared according to the invention is characterized in that the materials are conveniently taken and the ethical violation problem is avoided; the injection prepared according to the invention can effectively preserve the activity of stem cells, can be directly used for recovery, need not be cleaned and centrifuged and is conveniently used; after the injection prepared according to the invention is fed back, the differentiation of the mesenchymal stem cells to the hepatic cells and the maturation of the hepatic cells can be boosted and the GVHD occurrence rate can be reduced.

The invention discloses a full-suspension culture method for avian influenza virus. The full-suspension culture method comprises the following steps: step 1, taking EB66 cells for growth culture; step2, when the density of the EB66 cells grows to be suitable for virus inoculation, utilizing a fed-batch fluid-replacement virus inoculation technology to inoculate avian influenza virus into the EB66cells and performing virus multiplication culture; step 3, 24h after virus inoculation, sampling every 12h to determine HV titer of the virus and obtaining and storing the virus when the virus HA titer reaches to the maximum to obtain cultured avian influenza virus. According to the full-suspension culture method disclosed by the invention, the EB66 cells are utilized to perform avian influenza virus culture, so that the defect that a lot of miscellaneous protein is prone to being introduced into when a traditional chick embryo culture technology is utilized for production is overcome, and occurrence of chicken immunity side reaction is effectively reduced; meanwhile, titer and purity of the cultured avian influenza virus are improved; furthermore, production quality of the avian influenza vaccine is improved.

The invention discloses a preparation method of an immortalized human intervertebral disc nucleus pulposus cell system. The preparation method is as follows: extrinsic human telomerase reverse transcriptase is transfected into the human nucleus pulposus cell to activate telomerase and construct immortalized human nucleus pulposus cell line through induction immortalized human nucleus pulposus cell line, normal cells not transformed cells are used, wherein the cells have normal growth rate and karyotype, have contact inhibition, anchorage dependence and other safety properties and do not have carcinogenicity and soft agar cloning capability; and as same as the germline stem cells, the cells have real immortalization, thus the standard cell line can be provided for the further research of the pathogenic mechanism of the degenerative disc disease and a new idea is provided to increase the entire control level of the degenerative disc disease.

The invention relates to a medicine for curing osteonecrosis, belonging to the field of regenerative medicine engineering. Human amniotic mesenchymal cells (hAMCs) are used as cell source for cellular transplantation and tissue organization so as to cure the osteonecrosis and to provide a new way for repairing a damaged osseous tissue and reorganizing tissues. Based on the multi-lineage differentiation potential of the hAMCs, osteocytes, osseous tissues and vascular endotheliums can be differentiated in the human body, thereby completing the processes of osteanagenesis, bone remodeling and vascularization and forming a new osseous tissue integrated with the tissues of the patient.

The invention discloses a recombinant Marek's disease virus strain SCA13 strain. The biological preservation number of the strain is CGMCC NO.15285, and an SCA13 strain genome is integrated with ALV LTR and can exist stably. On the basis of BAC clone, and by means of the phenomenon that a Red E/T recombinant technology lacks meq and vTR genes, a deletion strain of the meq and vTR genes of the MDVSCA13 strain is built, the biological preservation number of the strain is CGMCC NO.15286, a double gene deletion strain built on the basis of the SCA13 strain completely loses lethality and tumorigenicity to chickens, the immunosuppressive property is lost, the strain has better biosecurity and better horizontal transmission capacity, the problem that in the MD vaccine immunity process, due to immunity leakage of the chickens, diseases occur can be solved, and the chickens are subjected to better immunoprotection in clinic.

The invention discloses an immortalized human neural stem cell line and a preparation method thereof. The method comprises the following steps: (1) isolating and culturing primary human hippocampus-derived neural stem cells; (2) constructing a recombinant lentiviral vector containing L-myc gene; (3) carrying out packaging production of the above recombinant lentiviral and collecting supernatant; (4) selecting virus supernatant with appropriate concentration for transfection and screening of stem cells and constructing the immortalized human neural stem cell line. The preparation method of immortalized cell line provided by the invention is a novel stable and safe and effective immortalized coding strategy; the prepared immortalized human neural stem cell line has all the biological characteristics of primary neural stem cells, and also has the feature of normal three-line differentiation at the same time, can be successfully differentiated into neurons, astrocytes and oligodendrocytes,and the growth rate of such cell line and ability to form neurospheres are stronger than the growth rate of primary cells, and the cell line can still proliferate rapidly after 20 generations withouttumorigenicity.

A preparation method of the adipose-derived stem cell exosome comprises the following steps: culturing 3rd to 6th generations of mouse adipose-derived stem cells in logarithmic growth by using a complete culture solution containing FBS until the cell density reaches 70 to 80 percent, and continuously culturing for 45 to 50 hours by using a DMEM culture solution containing exosome-free FBS, so as to obtain the adipose-derived stem cell exosome. And collecting a supernatant culture solution, centrifuging to remove cells and cell debris, centrifuging at 1500-2500g by using an ultrafiltration tube with the molecular weight of 100kDa for 30 minutes, concentrating, adding an exosome rapid extraction reagent into a concentrated solution, extracting, centrifuging, and collecting the mouse adipose-derived stem cell exosome. In-vitro and in-vivo experiments show that the mouse adipose-derived stem cell exosome can significantly promote hair growth and has an outstanding effect.

The invention discloses a method for preparing rabies vaccine for veterinary use. According to the method, the rabies vaccine for veterinary use is cultivated and produced by a cell bioreactor total-suspension non-serum method. In particular, the method comprises the following steps: performing virus infection on a BHK-21 cell with certain density and adapted to a strain by utilizing a rabies strain PV/BHK-21 and performing virus harvesting for many times, wherein the BHK-21 cell is obtained by performing total-suspension, non-serum and continuous perfusion cultivation by the cell bioreactor;and performing filtering system treatment and deactivation on the harvested virus stock solution, adding an adjuvant to prepare the rabies inactivated vaccine with high immunity and high safety. Through total-suspension and continuous perfusion culture by the cell bioreactor, high-density cell culture can be realized and viruses can be harvested continuously; and through non-serum culture, serum source pollution can be avoided, the purification process can be simplified and allergic reaction of the organism caused by the serum can be reduced. By the method provided by the invention, mass production and popularization can be realized.

A cell product for prevention and/or treatment of vascular disorders such as aortic aneurysm, comprising a SSEA-3-positive pluripotent stem cell derived from a mesenchymal tissue in a living body or a cultured mesenchymal cell (Muse cell).

The invention provides a culture medium and a method for inducing airway basal stem cells to differentiate into bronchoalveolar stem cells, and relates to the technical field of stem cell induced differentiation. The culture medium comprises a primary induced differentiation culture medium and a secondary induced differentiation culture medium; the primary induced differentiation culture medium comprises the following components: a fibroblast growth factor 10, a keratinocyte growth factor and a glycogen synthase kinase-3 inhibitor; the secondary induced differentiation culture medium is prepared from the following components: a fibroblast growth factor 10, a keratinocyte growth factor, a keratinocyte growth factor, a gamma secretase inhibitor and a phosphodiesterase inhibitor. According to the method disclosed by the invention, the BC is subjected to induction culture by adopting the induction culture medium and can be efficiently converted into the BASC. According to the culture medium and the culture method, autologous airway basal stem cells can be successfully induced into bronchoalveolar stem cells, and the induction conversion rate is high.

The invention relates to a method for constructing a cell model and cell bank for chromosomal translocation by recombination of hTERT (human Telomerase Reverse Transcriptase), and belongs to the field of medical science. The method is mainly characterized by comprising the following steps of carrying out dual-enzyme digestion on a plasmid pCIneo-hTERT and a vector pLXSNneo by virtue of endonucleases EcoR I and Xho I to obtain hTERT and pLXSNneo enzyme-digested products by linking through Ligation Mix, carrying out PCR amplification and separating through gel electrophoresis, constructing a pLXSNneo-hTERT recombinant, transforming the recombinant with DH5a competent cells, purifying, amplifying and picking ampicillin-resistant colonies, extracting a plasmid, transfecting with a lipidosome, carrying out in-vitro passage on chromosomal translocation cells in logarithmic growth so that the recombinants and cell DNA are integrated, carrying out enlarge culture, cloning a cell containing positive recombinants screened through G418, screening the cell of which a cell morphology, a growth curve, a chromosome karyotype, carcinogenicity tests of naked mice, the activity of telomerase in transfected cells, expression products of hTERT mRNA, immunohistochemical staining, a cell proliferate cycle and an apoptosis rate are in line with characteristics of an immortalized cell and are same as or similar to those of a primary cell as the hTERT-mediated cell model for chromosomal translocation, and cryopreserving in liquid nitrogen. The cell model lays a foundation for in-vitro research on the pathogenesis of chromosomal translocation in a cellular level in a long term.