34 results about "Oct-4" patented technology

A source of multipotent amniotic fluid / fetal stem cells (MAFSCs) is disclosed. MAFSC are of fetal origin and have a normal diploid karyotype. These cells are characterized by the following cell surface markers: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, HLA class I, CD13, CD44, CD49b, CD105 and are distinguished by the absence of the antigen markers CD34, CD45, and HLA Class II, but are distinguished from mouse embryonic stem cells in that these cells do not express the cell surface marker SSEA1. MAFSC express the stem cell transcription factor Oct-4. MAFSC cells can be propagated for an indefinite period of time in continuous culture in an undifferentiated state. The MAFSCs have the ability to differentiate in culture in a regulated manner, into three or more subphenotypes. Cells can then be differentiated into endodermal, mesodermal and ectodermal derived tissues in vitro and in vivo. A method for isolating, identifying, expanding and differentiating MAFSCs is disclosed.

Cells which may be differentiated at least into nerve cells, which are useful for therapies of brain metabolic diseases, are disclosed. The cells are side population cell separated from human amniotic mesenchymal cell layer, in which expressions of Oct-4 gene, Sox-2 gene and Rex-1 gene are observed by RT-PCR, and which are vimentin-positive and CK19-positive in immunocytostaining.

The present invention relates to methods for reprogramming a somatic cell to pluripotency by administering into the somatic cell at least one or a plurality of potency-determining factors. The invention also relates to pluripotent cell populations obtained using a reprogramming method.

Cells which may be differentiated at least into nerve cells, which are useful for therapies of brain metabolic diseases, are disclosed. The cells are side population cell separated from human amniotic mesenchymal cell layer, in which expressions of Oct-4 gene, Sox-2 gene and Rex-1 gene are observed by RT-PCR, and which are vimentin-positive and CK19-positive in immunocytostaining.

A method of harvesting mesenchymal stem cells from human amniotic fluid uses a two-stage culture protocol comprising culturing human amniocytes and then culturing mesenchymal stem cells. For culturing human amniocytes, primary amniocyte cultures are set up using routine or standard culture protocol in a cytogenetic laboratory. Non-adherent human amniotic fluids cells in the supernatant medium are collected. For culturing mesenchymal stem cells (“MSC”), the non-adherent cells are centrifuged and then plated with an alpha-modified Minimum Essential Medium supplemented with fetal bovine serum. Incubate with humidified CO2 for MSC growth. Reverse transcription polymerase chain reaction (“RT-PCR”) and immunocytochemical analyses reveal that Oct-4 mRNA and OCT-4 protein expression is detectable in the cultured amniotic fluid mesenchymal stem cells (“AFMSCs”). Under differentiation culture conditions, the AFMSCs can be induced to develop into multi-lineage cells, such as adipocytes, osteocytes, neuronal cells, etc.

The invention provides for isolated population of pulp marrow similar cells (DPMSCs) and methods for isolating and using these cells. The population of DPMSCs are highly homogenous for CD1O, CD29, CD13, CD44, CD49a, CD49d, CD59, CD73, CDw90, CD105, Oct-4 Isoform A and B, Nanog, Sox-2, and SSEA-4.

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 CD90, CD66A, CD66E, CXCR4, CD133 or an SSEA. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1, TWIST, KLF-4 and Stella but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105, CD90, CD66A, CD66E, CXCR4, CD133 or an SSEA. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiated the stem cells, are also provided.

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1 and TWIST but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiated the stem cells, are also provided.

The invention provides a novel application of icariin. Expression of a pluripotent gene Oct-4mRNA can be improved after the icariin acts on an umbilical cord mesenchymal stem cell after a week, pluripotency of the umbilical cord mesenchymal stem cell is improved, the capacity of the umbilical cord mesenchymal stem cell differentiating a nerve cell, a myocardial cell and an islet cell can be improved, the methylation level of the pluripotent adjusting gene Oct-4 is reduced, and the acetylation level of histone H3K9 is promoted.

Described herein are reprogramming techniques allowing for production of mammary-derived iPSCs (“m-iPSCs”). The m-iPSCs described herein exhibit all the hallmarks of stem cell identity including round cluster, bright colony morphology, clonal expansion, and pluripotent marker expression (alkaline phosphatase expression, Oct-4, nanog, etc.) Further refined techniques allow for generation of m-iPSCs under essentially defined conditions.

The invention relates to a method for extracting sub-totipotent stem cells from a chorion of a fetal surface of a placenta. The method comprises the steps of removing an amnion and stagnated blood from the placenta, and then repetitively washing the surface of the placenta to perform sterilization; shearing the chorion of the fetal surface in a glass utensil, removing placenta lobule tissues remained on the surface as much as possible, and shearing the chorion into small blocks; washing small tissue blocks by using a screen and a great amount of normal saline, and removing residual blood cells; performing tissue digestion: performing oscillatory digestion in a constant temperature shaker by using mixed enzymes; adding a proper amount of FBS for termination after digestion, performing filtration through a filter screen, and adding a great amount of normal saline to wash filter residues to obtain cells as many as possible; performing centrifugation, abandoning supernatant, adding saline water for washing and performing centrifugation to obtain mononuclear cells; performing magnetic bead sorting (OCT-4 positive, Nanog positive and STRO-1 negative) to obtain target cells. The invention further relates to the sub-totipotent stem cells obtained by adopting the method and pharmaceutical use thereof. The sub-totipotent stem cells have excellent characteristics.

Purified preparations of human embryonic stem cells with certain population-specific characteristics are disclosed. This preparation is characterized by the positive expression of the following pluripotent cell surface markers: SSEA-1 (−); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); alkaline phosphatase (+), as well as a set of ES cell markers including Oct-4, Nanog, Rex1, Sox2, Thy1, FGF4, ABCG2, Dppa5, UTF1, Cripto1, hTERT, Connexin-43 and Connexin-45. The cells of the preparation are negative for lineage specific markers like Keratin 8, Sox-1, NFH (ectoderm), MyoD, brachyury, cardiac-actin (mesoderm), HNF-3 beta, albumin, and PDX1 (endoderm). The cells of the preparation are human embryonic stem cells, have normal karyotypes, exhibit high telomerase activity and continue to proliferate in an undifferentiated state after continuous culture for over 40 passages. The embryonic stem cell line Relicell™ hES1 also retains the ability, throughout the culture, to differentiate into cell and tissue types derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). Methods for isolating a human embryonic stem cell line are also disclosed.

The invention relates to the technical field of medical biology engineering, and provides an anti-human Oct 4 (octamer-binding protein 4) monoclonal antibody, recombinant protein for preparing the monoclonal antibody, a hybridoma cell strain secreting the monoclonal antibody as well as applications of the monoclonal antibody in western blot, expression of Oct 4 in immunohistochemical detection samples and preparation of prognosis evaluation preparations for liver cancer.

The present invention relates to multipotent adult stem cells expressing Oct4, derived from umbilical cord blood (UCB) and also these cell are expressing CD29, CD31, CD44, simultaneously, a method for preparing the same, and more specifically to multipotent adult stem cells which are obtained by culturing umbilical cord blood-derived monocytes in a medium containing bFGF (basic fibroblast growth factor) and human serum or plasma. In addition, multipotent adult stem cells expressing Oct-4 from UCB are morphologically spindle or round shaped cells Although the stem cells according to the present invention are adult stem cells, they are multipotent and capable of differentiating into ectodermal-, messodermal-, endodermal-originated tissue or cells including osteogenic cells or nerve cells etc, thus they can be effectively used in the treatment of intractable diseases and incurable diseases.

The invention belongs to the field of pharmacy and relates to application of baicalein as an SHH (Sonic Hedgehog) signaling pathway inhibitor in the preparation of drugs for treating pancreatic cancer; results of animal experiments show that baicalein regulates Sonic Hedgehog signaling pathway to inhibit the proliferation of pancreatic cancer cells, induce apoptosis, and inhibit self-update of pancreatic cancer stem cells; the baicalein down-regulates IC50 of PANC-1 pancreatic cancer cells, weakening the in-vitro motor ability of the cells; the baicalein down-regulates the activity of CD44+ / CD24+PANC-1 pancreatic cancer cells and down-regulates surface marker CD24 and CD44 protein expression for pancreatic cancer cells, thereby reducing nuclear transcription factor OCT-4 and SOX-2 protein-level expression, inhibiting SHH, PTCH-1, SMO and GLI2, and reducing OCT-4 and SOX-2 protein-level expression. The baicalein may act as an SHH signaling pathway inhibitor to prepare drugs for treating pancreatic cancer.

The invention discloses a method for separating and purifying totipotent stem cells from the tissues of early aborted fetuses of human beings, including the steps: a. a sample of the placenta tissue of the aborted fetus after the first pregnancy period is collected; b. the umbilical cord tissue or the intestine tissue of the fetus is obtained from the sample gained in the step a; c. cells are obtained from the umbilical cord tissue or the intestine tissue of the fetus gained in the step b; and d. markers of the embryonic stem cell, such as OCT-4, Nanog, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, and the like which are obtained from the cells of the umbilical cord tissue or the intestine tissue of the fetus, are separated, purified and expressed, and can form the stem cell of an embryoid body. The cells obtained by the method of the invention are always in the state of undifferentiation. The invention also discloses the application of the stem cells in forming different human cells, such as cardiac muscles, nerves and islet beta cells or osseus cells. The method avoids the dispute over ethnics which needs to be faced with during the operation of obtaining the totipotent stem cells from the embryo, and more stem cells can be obtained than from blastocyst.

The present invention relates to the production of avian induced pluripotent stem cells from non-pluripotent somatic cells, including embryonic fibroblasts and adult somatic cells. In this method, avian (including quail or chicken) somatic cells are reprogrammed into a state closely resembling embryonic stem cells including the expression of key stem cell markers alkaline phosphatase, etc. by transfecting / transducing the non-stem cells with genes (preferably using a non-integrating vector as otherwise described herein or alternatively an integrating vector, such a lentiviral vector, retroviral vector or inducible lentiviral vector, among others) which express at least nanog, Lin28 and cMyc. In preferred aspects of the invention, the transfected / transduced vectors express nanog, Lig28, cMyc, Oct 4 (POU5F1 or PouV), SOX2 and KLF4. The induced stem cells which are produced contribute to all 3 germ layers, the trophectoderm and in certain aspects, the gonad in chimeric offspring.

The present invention discloses a novel population of multipotent cardiac precursor (MCP) cells derived from human blastocysts derived stem cells, methods for the preparation thereof and use of the cells for in vitro testing. Basement cells derived from hBS cells are also disclosed and method for the preparation of MCP cells from basement cells. The MCP cells have the following characteristics i) at least 1 % of the cells exhibit no antigen expression of one or more markers for undifferentiated cell, the marker being selected from the group consisting of SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and Oct-4, ii) at least 1% of the cells exhibit no protein expression of one or more of a neural marker including nestin or GFAP iii) at least 1% of the cells exhibit protein and / or gene expression of one or more of a mesodermal marker including brachyury, vimentin or desmin iv) at least 1% of the cells exhibit protein and / or gene expression of Flk-1 (KDR). Furthermore, the MCP cells have a characteristic morphology. They grow as clusters of small, round and phase-bright cells; individual cells are 5-20 [mu]m in diameter and each cluster is composed of 2-500 cells. They form clusters of round or elongated shape, that appear as loosely adherent cell clumps that as illustrated in figure 2 panel a, b and c. Furthermore, they have a relatively high nucleus-to-cytoplasma ratio, e.g. 1 :2-1 :64 of the total volume of the cell and / or appear as balloons on a string, as illustrated in figure 18, schematic sketch. Moreover, the MCP cells are non-contracting.

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1 and TWIST but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiated the stem cells, are also provided.

The invention provides a method for inhibiting PUMA function and improving iPS (induced Pluripotent Stem) cell construction effect. The method comprises the following steps of: (1) preparing fibroblast MEF (Mouse Embryonic Fibroblast) of a PUMA- / - (PUMA gene knocked out) mice or PUMA- / + mice, and performing passage and frozen storage; (2) converting plasmid PMX-Oct 4, PMX-Sox2, PMX-c-Myc and PMX-KLF4 through DH5 alpha escherichia coli strain, and respectively highly and slightly extracting plasma; (3) packing retrovirus, tittering and performing frozen storage; (4) knocking out reprogramming of embryo fibroblasts MEF of the mice or PUMA- / + mice through PUMA gene; and (5) knocking down the programming of human fibroblast of the PUMPA function. The method has the beneficial effects that the PUMA function is inhibited and the iPS cell construction effect is improved during programming somatic cell; the stability of iPS cell genome can be protected at the same time.

The invention discloses a method for separating and purifying human amniotic fluid stem cells. The method is to use an embryo antigen-4 at a special stage as a screening marker to separate and purify the human amniotic fluid stem cells which have homogeneous characteristics and are positively expressed by the embryo antigen-4 at the special stage from the human amniotic fluid stem cells which are adherently cultured. Experimental results show that the method can obtain the human amniotic fluid stem cells which have homogeneous characteristics, good capacities of proliferation and inductive differentiation, and cell ratio of G0 stage to G1 stage cells of more than 90 percent, and can express specificity markers of human totipotent stem cells such as the embryo antigen-4, Nanog and Oct-4 at the special stage. The invention provides a new way to obtain the human amniotic fluid stem cells, and lays a foundation for the further application of the human amniotic fluid stem cells.

The invention relates to a method for extracting sub-totipotent stem cells from a chorion of a fetal surface of a placenta. The method comprises the steps of removing an amnion and stagnated blood from the placenta, and then repetitively washing the surface of the placenta to perform sterilization; shearing the chorion of the fetal surface in a glass utensil, removing placenta lobule tissues remained on the surface as much as possible, and shearing the chorion into small blocks; washing small tissue blocks by using a screen and a great amount of normal saline, and removing residual blood cells; performing tissue digestion: performing oscillatory digestion in a constant temperature shaker by using mixed enzymes; adding a proper amount of FBS for termination after digestion, performing filtration through a filter screen, and adding a great amount of normal saline to wash filter residues to obtain cells as many as possible; performing centrifugation, abandoning supernatant, adding saline water for washing and performing centrifugation to obtain mononuclear cells; performing magnetic bead sorting (OCT-4 positive, Nanog positive and STRO-1 negative) to obtain target cells. The invention further relates to the sub-totipotent stem cells obtained by adopting the method and pharmaceutical use thereof. The sub-totipotent stem cells have excellent characteristics.

The invention relates to a pair of human embryonic stem cell lines with the same HLA property. The paired human embryonic stem cell lines have the same HLA property, normal karyograms and higher affinities for expressing specific surface antigens; the paired human embryonic stem cell lines are strongly positive through AKP detection, and simultaneously, can express Oct-4 transcription factors and display undifferentiated properties; and the average population doubling time is about 32 hours. More choices for solving HLA mating difficulty are provided by researching the properties of the pair of human embryonic stem cell lines from the same male parent and the same female parent in biology, genetics and epigenetics, and meanwhile, the two embryonic stem cell strains provide good models for clinical basic research.

The present invention relates to a novel method for preparing induced pluripotent stem cells (iPSCs) by introducing four genes, Oct-4, Sox2, Klf4, and Glial, into somatic cells. The present invention also relates to the iPSCs produced by the aforementioned method. Also provided is a process of drug selection for a heritable genetic disease by use of the iPSCs produced by the aforementioned method. In particular, wherein the inherited disease is Fabry disease. The present invention also relates to a method for treating Fabry-associated myocardiopathy in a subject in need thereof, and a method for determining prognosis in a subject with Fabry-associated myocardiopathy.

The invention belongs to the field of cells, and discloses a method for inducing fetal bovine fibroblast to reprogram. According to the invention, the fetal bovine fibroblast is subject to permeabilized treatment by a digitonin solution, and is subject to co-incubation with xenopus oocytes extract; whole medium containing 2mMcaCl2 is adopted to block reversible pores in the cell membrane, extract treated medium is adopted to cultivate and induce the fetal bovine fibroblast to reprogram to obtain fetal bovine fibroblast pluripotent stem cells. The method discloses by the invention is efficient, stable, and safe; the alkaline phosphatase staining of the fetal bovine fibroblast pluripotent stem cells disclosed by the invention shows a positive result; the acetylation degree of histone H3K9 is lowered; Oct-4 protein and pluripotent marker gene Oct-4, Nanog are expressed; therefore the fetal bovine fibroblast undergoes reprogramming, the gene expression mode of cells is changed to show characteristics of stem cell, partial growing totipotent is restored, and the fetal bovine fibroblast can be used as nuclear donor cells to prepare clone embryo for body cell clone.

The present invention provides a method for treating leukemia utilizing somatic cell reprogramming. The method includes a step of introduction of somatic cell reprogramming inducing factors Oct-4, Sox-2, Klf4 and c-Myc (OSKM for short) into leukemic cells or a step of utilizing small reprogramming molecules in in-vitro culture. It promotes leukemic cells to initiate process of somatic cell reprogramming in order to induce apoptosis and finally purpose of eliminating leukemic cells in-vivo or in-vitro is achieved. It provides new ideas and methods for clinical treatment of leukemia in the future.

The present invention relates to multipotent adult stem cells expressing Oct4, derived from umbilical cord blood (UCB) and also these cell are expressing CD29, CD31, CD44, simultaneously, a method for preparing the same, and more specifically to multipotent adult stem cells which are obtained by culturing umbilical cord blood-derived monocytes in a medium containing bFGF (basic fibroblast growth factor) and human serum or plasma. In addition, multipotent adult stem cells expressing Oct-4 from UCB are morphologically spindle or round shaped cells Although the stem cells according to the present invention are adult stem cells, they are multipotent and capable of differentiating into ectodermal-, messodermal-, emdodermal-originated tissue or cells including osteogenic cells or nerve cells etc, thus they can be effectively used in the treatment of intractable diseases and incurable diseases.

Retinal stem cells haying embryonic-like characteristics are disclosed. The retinal stem cells express one or more of the embryonic stem cell markers Nanog and OCT-4. The retinal stem cells may be obtained from retinal pigment epithelium. Methods of making and using retinal stem cells are also disclosed.

The present invention relates to a cancer initiating cell comprising an isolated coxsackievirus and adenovirus receptor positive mouse pulmonary stem / progenitor cell that overexpresses Oct-4.