134 results about "Stem cell line" patented technology

Methods are disclosed for generating HLA homozygous parthenogenetic human stem cell (hpSC-Hhom) lines from both HLA homozygous and HLA heterozygous donors. These hpSC-Hhom lines demonstrate typical human embryonic stem cell morphology, expressing appropriate stem cell markers and possessing high levels of alkaline phosphatase and telomerase activity. Additionally, injection of these cell lines into immunodeficient animals leads to teratoma formation. Furthermore, in the case of HLA heterozygous donors, the hpSC-Hhom lines inherit the haplotype from only one of the donor's parents. SNP data analysis suggests that hpSC-Hhom lines derived from HLA heterozygous oocyte donors are homozygous throughout the genome as assessed by single-nucleotide polymorphism (SNP) analysis. The protocol as disclosed minimizes the use of animal-derived components, which makes the stem cells more practical for clinical application.

A method of establishing a clonal embryonic stem cell line capable of sustaining a phenotype of normal embryonic stem cells following at least eight months of in vitro culture is disclosed. The method is effected by culturing an individual embryonic stem cell for at least eight months in a serum-free medium, thereby establishing the clonal embryonic stem cell line capable of sustaining said phenotype of normal embryonic stem cells following at least eight months of in vitro culture.

This present invention provides novel methods for deriving embryonic stem cells, those cells and cell lines, and the use of the cells for therapeutic and research purposes without the destruction of the embryo. It also relates to novel methods of establishing and storing an autologous stem cell line prior to implantation of an embryo, e.g., in conjunction with reproductive therapies such as IVF.

A method for isolating an inner cell mass comprising the steps of immobilizing a blastocyst stage embryo having a zona pellucida, trophectoderm, and inner cell mass, creating an aperture in the blastocyst stage embryo by laser ablation, and removing the inner cell mass from the blastocyst stage embryo through the aperture. The aperture is through the zona pellucida and the trophectoderm. The laser ablation is acheived using a non-contact diode laser. The inner cell mass removed from the blastocyst stage embryo is used to establish human Embryonic Stem Cell lines.

The invention discloses a dendrobium officinale stem cell and an isolated culture method thereof. The method comprises the following steps: culturing plant root tissue containing a quiescent centre, performing subcultring and scale-up culture on quiescent centre stem cell, and then preparing stem cell dry powder or extracting solution according to requirements. The dendrobium officinale stem cell overcomes the problem that dendrobium officinale in vitro material is easy to brown, the cell of a stem cell line is not differentiated, the cell increment is large in a nutritional condition, the culture time is short, and the cell activity is high. The dendrobium officinale stem cell and the method have the advantages that the technical process is easy to operate, the quantity of technological processes is less, the output is high, and the dendrobium officinale stem cell obtained is anti-aging and oxidation resistant, promotes hair growth and high baldness-resistant activity, and can be manufactured into functional health care products and cosmetics for preventing and curing various senescence and baldness and relevant Chinese drugs for preventing senescence and baldness.

The present invention provides methods for isolating individual viable stem cells from a stem cell line, methods for deriving one or more clones from a stem cell line, individual viable cells and clones derived from stem cell lines by the methods disclosed, methods for producing differentiated cells from the individual viable cells and clones so derived, differentiated cells so produced, methods for treating diseases using the cells and clones described herein and methods for proliferating cells and clones in undifferentiated form.

The present invention relates to the discovery that renewable stem cell lines can be derived from tumor cells and can be cultured in vitro. Accordingly, the invention provides neural tumor stem cell lines and cells from such cell lines. Because the cell lines retain characteristics of the tumors from which they are derived, the cells can be used in screening methods for identification of potential therapeutic agents and can be used to identify genetic markers which may be predictive for development of such tumors. Finally, such cells can be used to determine an appropriate therapeutic regimen for a patient suffering from a brain tumor. Cells from a patient's brain tumor can be cultured as described herein to create a cell line, and the relative effectiveness of a therapeutic agent against the cells can be tested to determine which agent or combination of agents is most effective in treating the patient's tumor.

The invention discloses a method for high-efficiency establishment of a genetically modified animal model through haploid stem cells. Specifically, the method for high-efficiency establishment of the genetically modified animal model through haploid stem cells comprises the following steps: a) knocking out the H19 gene of male haploid embryonic stem cells through homologous recombination; b) screening and identifying the male haploid embryonic stem cells in which the H19 gene is correctly knocked out; and c) injecting the male haploid embryonic stem cells in which the H19 gene is correctly knocked out into cytoplasm of oocyte so as to produce a semi-cloned genetically modified animal. The method provided by the invention simulates the functions of sperms by establishing an H19 gene-knocked-out male haploid embryonic stem cell system so as to highly efficiently generate fertile progeny, which can then be used for implementation of novel genetic manipulation and establishment of corresponding genetically modified animal models.

The invention discloses an immortalized porcine pancreatic stem cell line and construction and differentiation methods thereof. In the immortalized porcine pancreatic stem cell line, porcine pancreatic stem cells are taken as host cells and are transfected with a pCI-neo-hTERT eukaryotic expression vector, and the human telomerase reverse transcriptase screened by G418 is expressed and has a transformant with multi-directional differentiation potentiality. The cell line also has higher activity after transferring for over 50 generations in vitro, keeps split proliferation, does not generate aging or apoptosis, and is an immortalized cell line. The cell line can be differentiated to form functional islet cell mass after induction, can reduce the blood glucose concentration after being transplanted into a mouse model suffering from diabetes and can maintain a normal blood glucose level within two weeks.

The invention discloses an immortalized canine adipic mesenchymal stem cell line and a constructing method thereof. The immortalized canine adipic mesenchymal stem cell line capable of expressing SV40 large T antigen genes and having multidirectional differentiation is obtained by: transfecting a lentiviral vector carrying SV40 large T antigen (Ttag) by using canine adipic mesenchymal stem cells as host cells, integrating the Tag genes into an adipic mesenchymal stem cell genome and carrying out continuous passage and screening. This cell line is still active in case of 50 in-vitro passages, is high in differentiation and proliferation performance, never ages or dies, can also save medium cost and is an immortalized cell line.

The invention provides a human tumor stem cell line T3A-A3, which is obtained by separation of liver cancer tissue excised in an operation of a primary hepatic carcinoma patient. The separated cells can be in long-term passage culture in vitro, can grow rapidly and retain stem cell property after more than 100 times of passage. The cell can express markers of multiple stem cells, has self-updatingcapability of stem cells and directional differentiation potentials for different tumor cells, and also has tumor property, tumorigenicity ability and metastatic ability. The invention is a powerfultool for preparing stem cell drugs for targeting tumor, due to the facts that the cells can be in long-term passage culture in vitro and retain unchanged stem cell property, and the tumorigenicity ability and metastatic ability thereof are strong in immunodeficient mice.

The invention relates to a human parthenogenetic embryo stem cell line with two active X chromosomes and derivatives thereof. A karyotype of the human parthenogenetic embryo stem cell line is 46, XX before 10 generations; the karyotype of the cell line becomes a chimera from the 20th generation; all cells of the cell line lose an active X chromosome at the 35th generation; and in the inactivation detection of the X chromosome, only the state of the active X chromosome is maintained all along, but the presence of the inactivated X chromosome is not discovered. The parthenogenetic activation can be achieved by adopting an artificial activation condition causing a second polar body to be discharged; therefore, the human parthenogenetic embryo stem cell line and the derivatives thereof can be obtained. The cell line and the derivatives thereof have important significance on researching the characteristics of the cell on genetics and epigenetics and further applying the cell to clinically treating certain diseases; and a cell source for cell replacement therapy can be obtained by transforming genes of the cell or modifying the genes on the epigenetics.

Provided are an androgenetic haploid stem cell line, preparation method and use thereof. Specifically, provided are an androgenetic haploid cell and androgenetic blastula. The nucleus of the cell or the blastula only comprises a haploid autosome and a sex chromosome, the sex chromosome being the X chromosome and containing no Y chromosome. The androgenetic haploid cell of the present application can replace a spermatid as a ligand to generate a fertile animal individual, thus facilitating gene manipulation and being capable of transferring genetic information to the offspring.

Methods of developing and using cell lines, such as stem cell lines, for therapeutic or cosmetic use. In one embodiment the cell lines are used to treat a wide range of degenerative and metabolic disorders including, but not limited to, obesity, diabetes, hypertension, and cardiac deficiency. Also described are methods of using such cell lines to screen for compounds that play a role in regulating a variety of processes.

The invention provides a method for establishing a human nasopharyngeal carcinoma tumor stem cell line. The method comprises the following steps: (1) obtaining human nasopharyngeal carcinoma cells in the single cell state; (2) adding the human nasopharyngeal carcinoma cells in the single cell state, obtained in the step (1), to a serum-free stem cell culture medium containing epidermal growth factors and basic fibroblast growth factors to obtain single cell suspension; (3) adjusting the concentration of the single cell suspension in the step (2) to 900-1100 single cells/ml; (4) inoculating 1ml of the single cell suspension obtained in the step (3) into single pores on a cell culture six-pore plate and then adding the equivoluminal stem cell culture medium for culture for 72-96 hours; and (5) collecting the cells obtained in the step (4), carrying out trypsinization till the single cell and carrying out subculture. The method provided by the invention is simple and convenient to operate; and the obtained nasopharyngeal carcinoma stem cell line has obvious stem cell characteristics.

Production of secretory insulin dry cell system by encoded various exogenous gene slow virus carrier is carried out by copying pWPST, cloning various insulin regulating genes to the carrier, transfecting 293T cell, packing, collecting virus grains, concentrating and transfecting human embryo pancreas dry cell. It's simple, efficient and practical, it can obtain one, two or three kinds of exogenous gene cells, which are in different expression and meet different needs.

Two cell lines, PICM-19H and PICM-19B, were derived from the bipotent ARS-PICM-19 pig liver stem cell line and assessed for their potential application in artificial liver devices. The study included assessments of growth rate and cell density in culture, morphological features, and hepatocyte detoxification functions, i.e., inducible CYP450 activity, ammonia clearance, and urea production. The PICM-19H cells contain numerous mitochondria, Golgi apparatus, smooth and rough endoplasmic reticulum, vesicular bodies and occasional lipid vacuoles. PICM-19H cells display inducible CYP450 activity, clear ammonia, and produce urea in a glutamine-free medium. Ultrastructural analysis of the PICM-19B monolayers show that the roughly cuboidal cells display basal-apical polarization and are joined by tight junction-like complexes. Other ultrastructure features are similar to those of PICM-19H cells except that they possess numerous cell bodies resembling mucus vacuoles. The PICM-19B cells possess relatively high levels of GGT activity, but retain some inducible CYP450 activity, and some ammonia clearance and urea synthesis ability. These data indicate that both cell lines, either together or alone, may be useful as the cellular substrate for an artificial liver device. In vitro models of the liver are needed to replace animal models for the rapid assessment of drug biotransformation and toxicity. A unipotent porcine stem cell line PICM-19H differentiates exclusively into hepatocytes and can be induced to express CYP450 enzymes. These cells have many activities associated with xenobiotic phase I and phase II metabolism lacking in other liver cell lines. The PICM-19H cell line was also compared to the tumor-derived human HepG2 C3A cell line and to primary cultures of adult porcine hepatocytes. The results demonstrate the potential for the use of PICM-19H cells in drug biotransformation and toxicity testing and further support their use in artificial liver device technology.

The invention acquires an immortalized human umbilical cord mesenchymal stem cell line. Through the construction of a recombinant lentiviral plvx-hteret vector, a catalytic subunit hTERT of human telomerase is introduced into umbilical cord mesenchymal stem cells, so as to screen out a stem cell line capable of stably expressing hTERT. According to cell culture and identifications at various levels, it is proved that an immortalized umbilical cord mesenchymal stem cell line is obtained, and the cell line maintains the capacity of differentiation. The invention also confirms biological activity and function of the cell line through a variety of methods, and it is demonstrated that the cell line can be used as an effective vector for gene therapy. Through a mouse tumorigenicity experiment, the invention also indicates that the cell line is safe and has no tumorigenic tendencies.

The present invention generally relates to neural stem cells, preferably fetal neural stem cells and their progeny thereof. The present invention provides methods of isolating culturing and propagating neural stem cells and the development of neural stem cell lines and lineages. The present invention also relates to the use of neural stem cells and somatic cells and cells expressing the telomerase catalytic component (TERT) for gene targeting and gene knockout experiments and for producing genetically modified animals.